FTA Bacterial DNA Whatman

FTA Bacterial DNA Whatman Raccolta, trasferimento, stoccaggio e purificazione di DNA batterico La tecnologia FTA della Whatman offre un valido e semplice metodo per la preparazione di campioni per analisi genetiche da batteri. Occorre semplicemente applicare campioni di colture o campioni clinici sulla matrice FTA il DNA viene catturato e stabilizzato all istante, consentendone lo stoccaggio a temperatura ambiente, da analizzare al momento più opportuno. Gli agenti patogeni vengono inattivati all istante. Trasporto sicuro e purificazione in 30 minuti rendono la tecnologia FTA un indispensabile strumento di ricerca. Caratteristiche Principali Indicato per un ampia varietà di batteri Il DNA genomico pronto per la PCR può essere isolato rapidamente da una varietà di batteri gram-negativi e grampositivi, batteri spore-formanti e acido-resistenti senza alcun o minimo pretrattamento. Raccolta semplice Depositare colture o campioni clinici direttamente sulla matrice FTA (FTA Card Indicatrice). Perfetto per il vostro metodo FTA funziona, qualsiasi il metodo scelto per l identificazione di batteri: campioni da colture pure di batteri o campioni clinici. Temperatura ambiente Gli acidi nucleici vengono automaticamente stabilizzati senza la necessità di refrigerazione. Applicazioni Applicazioni diagnostiche Analisi biologiche Analisi di alimenti Analisi ambientali Rapida purificazione Il DNA viene purificato sulla FTA Card in tre semplici steps, in un singolo tubo, a temperatura ambiente. Il DNA rimane immobilizzato sulla matrice ed è subito pronto per la PCR. Se è necessario avere DNA in soluzione si può usare Whatman GenSpin. Manipolazione e trasferimento sicuri FTA inattiva gli agenti patogeni potenzialmente nocivi. Automatizzazione L utilizzo di sistemi automatici accelera le fasi di preparazione di diversi dischi prelevati contemporaneamente, ottimizza le risorse disponibili e standardizza la purificazione di DNA. I dischi prelevati dalla FTA Card possono essere sottoposti alle fasi di lavaggio e preparazione per la PCR su una diversi tipi di strumenti presenti in commercio (liquid handling robot). Monitoraggio acqua/aria Identificazioni molecolari Applicazioni di ricerca Tre semplici passaggi operativi per ottenere un DNA puro Dispensare il campione Purificare Prelevare un piccolo disco (punch) dal Campione

PROTOCOLLO APPLICATIVO DEL FTA BACTERICAL DNA Dispensare il campione Dispensare il campione sulla FTA Card. Lasciarlo asciugare completamente. Se si utilizza una Card Indicatrice, l area coperta dal campione cambierà colore da rosa a bianco indicando la posizione del campione stesso. Prelevare un piccolo disco Prelevare (mediante perforazione) un piccolo disco dal campione depositato sulla FTA Card. Purificazione con Reagente di Purificazione FTA (Purification Reagent FTA) Mettere il disco di campione in un tubo per PCR ed effettuare tre lavaggi con il Reagente di Purificazione FTA. Eliminare la soluzione utilizzata dopo ciascun lavaggio. Lavaggio con soluzione tampone TE-1 Effettuare due lavaggi con la soluzione tampone TE-1 (10mM Tris, 0,1 mm EDTA, ph 8,0). Eliminare la soluzione usata dopo ciascun lavaggio. Essiccazione Lasciar asciugare il disco nel tubo PCR PCR Aggiungere la master mix per la PCR direttamente al disco e iniziare la reazione di amplificazione. Qualità Whatman Whatman, azienda leader nelle tecnologie per la separazione, è nota nellà comunità scientifica per prodotti e soluzioni innovative contraddistinte da un elevata qualità. Il nostro istinto per la semplificazione accelera la velocità di fare nuove scoperte, riduce i costi e risparmia tempo. Inoltre, per accentuare ulteriormente l impegno nel soddisfare le specifiche esigenze dei nostri clienti, Whatman è organizzata in quattro aree distinte: LabScience, BioScience, MedTech Diagnostica e MedTech Dispoitivi. Ulteriori informazioni sono disponibili collegandovi al sito www.whatman.com. FTA, GenSpin e Whatman sono marchi registrati della Whatman Group. INFORMAZIONI PER L ORDINE DAL CATALOGO WHATMAN No. Cat. Descrizione Quantità per Confezione WB120061 FTA Starter Pack 1 WB120204 FTA Reagente di Purificazione 500mL WB120205 FTA Classic Card (non-indicatrice) 100 WB120206 FTA Classic Card (indicatrice) 100 WB120055 FTA Mini Card (non-indicatrice) 100 WB120056 FTA Mini Card (indicatrice) 100 WB120210 FTA Micro Card (non-indicatrice) 100 WB120211 FTA Micro Card (indicatrice) 100 WB100005 Harris Micro Perforatore 1.2mm 1 WB100006 Harris Micro Perforatore 1.2mm Tip 1 WB100028 Harris Uni-Core 1.25mm Perforatore 4 WB100010 Busta Multi-Barrier (grande) 500 WB100011 Busta Multi-Barrier (piccola) 500 WB100025 1.2mm Plunger di Ricambio 1 WB100003 Essiccante (1g) 1000 WB100016 Busta Postale per FTA Card 50 WB100020 Tagliere di Ricambio 1 WB120005 GenSpin Kit di Purificazione DNA 50 Purificazioni Purificazione del DNA in meno di 25 minuti! Il GenSpin Kit di Purificazione del DNA della Whatman rappresenta un metodo estremamente semplice per purificare il DNA a singolo filamento da campioni di sangue e cellule in coltura. Il campione è subito pronto per la PCR a partire anche da piccoli volumi del campione di partenza. North America Europe Japan Asia Pacific Whatman Inc 9 Bridewell Place, Clifton, NJ 07014 Technical Support: 1-800-922-0361 Customer Service: 1-800-631-7290 Outside US: 973-773-5800 Fax: 973-773-0168 E-mail: info@whatman.com Whatman International Ltd Springfield Mill James Whatman Way Maidstone Kent ME14 2LE, UK Tel: +44 (0)1622 676670 Fax: +44 (0)1622 691425 E-mail: information@whatman.com Whatman Japan KK Daiwa Ueno Building 1F 6-10 Ueno 5-chome, Taito-ku Tokyo 110-0005, Japan Tel: +81 (0)3 3832 6707 Fax: +81 (0)3 3832 6457 E-mail: japaninfo@whatman.com Whatman Asia Pacific Pte Ltd 171 Chin Swee Road #08-01 San Centre Singapore 169877 Tel: +65 6534 0138 Fax: +65 6534 2166 E-mail: wap@whatman.com GenSpin TM Kit di Purificazione DNA Whatman Leaders in Separations Technology www.whatman.com Cat No. S9036-782

FTA Cards and Indicating FTA Cards WHATMAN CATALOGUE ORDERING INFORMATION Catalogue Number WB120205 WB120206 WB120055 WB120056 WB120210 WB120211 WB120208 WB120065 WB120028 FTA Classic Card Indicating FTA Classic Card* FTA Mini Card Indicating FTA Mini Card* FTA Micro Card Indicating FTA Micro Card* FTA Gene Card** PlantSaver Card CloneSaver Card * Indicating FTA Cards change colour from pink to white when sample is applied and are recommended for use with clear samples. **FTA Gene Cards are compatible with automated liquid sampling systems when used with FTA Gene Card Trays. FTA Reagent, Accessories and Kits WHATMAN CATALOGUE ORDERING INFORMATION Catalogue Number WB100016 WB100030 WB120061 WB120067 WB120204 WB100032 WB100003 WB100011 WB100010 WB100024 WB120052 WB100005 Description Cards/ Pack 100 100 100 100 100 100 100 100 5 Description Sample Areas/ Card FTA Card Mailer FTA Gene Card Tray FTA Starter Pack FTA Kit FTA Purification Reagent Sterile Foam Tipped Applicator Swabs Desiccant (1gm) Multi-Barrier Pouch, Small (for Mini, Micro and Gene Cards) Multi-Barrier Pouch, Large (for Classic Cards) CloneSaver Resealable Multi-Barrier Pouch CloneSaver Starter Kit Harris Micro Punch 1.2mm (with Mat) 4 4 2 2 1 1 3 4 96 Maximium Volume/ Sample Area (µl) 125 125 125 125 125 125 75 N/A 5 Maximium Total Volume/Card (µl) 500 500 250 250 125 125 225 N/A 480 Qty per Pack 50 20 1 1 500mL 100 1000 500 500 50 1 1 Whatman Quality Whatman is a global leader in separations technology and is known in the scientific community for providing innovative Life Science products and solutions. Our instinct for simplification accelerates the rate of discovery, reduces costs and saves time. In order to focus on the unique needs of our customers, Whatman is organised into four business development units: Analytical Chemistry, Diagnostics, Genomics & Proteomics and Medical Devices. For more information, visit www.whatman.com. CloneSaver, FTA, PlantSaver and Whatman are registered trademarks of the Whatman Group. North America Europe Japan Asia Pacific Whatman Inc. 200 Park Avenue Florham Park, NJ 07932 USA Technical Support: 1-800-922-0361 Customer Service: 1-800-631-7290 E-mail: info@whatman.com Whatman International Ltd Springfield Mill James Whatman Way, Maidstone Kent ME14 2LE UK Tel: + 44 (0)1622 676670 Fax: + 44 (0)1622 691425 E-mail: information@whatman.com Whatman Japan KK Daiwa Ueno Building 1F 6-10 Ueno 5-chome, Taito-ku Tokyo 110-0005, Japan Tel: +81 (0)3 3832 6707 Fax: +81 (0)3 3832 6457 E-mail: japaninfo@whatman.com Whatman Asia Pacific Pte Ltd 171 Chin Swee Road #08-01 San Centre Singapore 169877 Tel: +65 6534 0138 Fax: +65 6534 2166 E-mail: wap@whatman.com WB100028 Harris Uni-Core Disposable 1.25mm Punch (with Mat) 4 WB100007 Harris Micro Punch 2.0mm (with Mat) 1 WB100029 WB100020 Harris Uni-Core Disposable 2.0mm Punch (with Mat) Replacement Cutting Mat 4 1 Leaders in Separations Technology www.whatman.com WB100006 Replacement Tip 1.2mm 1 WB100008 Replacement Tip 2.0mm 1 Cat No. S9036-756

Whatman FTA Collect, archive, transport and purify nucleic acids all at room temperature Whether you re in a laboratory or deep in a rain forest, Whatman FTA provides a remarkably easy way to collect and isolate nucleic acid samples for analysis. Simply apply virtually any type of biological sample to the FTA matrix, and the nucleic acids are instantly captured and stabilised. Pathogens are inactivated, making samples safe to handle and ship. Store samples, including clones, at room temperature and analyse whenever you re ready. Try FTA, and you ll soon find it s an indispensable part of your nucleic acids toolbox. Features and Benefits Three easy steps to pure nucleic acids Simple collection Protection of nucleic acids from degradation at room temperature allows for convenient collection in the laboratory or the field. Room temperature storage Nucleic acids are automatically stabilised without the need for refrigeration. Pathogen inactivation Cells are automatically lysed on contact with the FTA matrix. Pathogens become inactivated, making samples safe to handle and ship via standard mail. Fast purification Nucleic acids are purified on the FTA Card in three simple steps, all in a single tube at room temperature. DNA remains immobilised on the matrix and is ready for PCR or other amplification techniques. Automatable Automation speeds the handling of multiple FTA punches and standardises DNA purification. Punches can be easily washed and prepared for PCR on a variety of liquid handling instruments. Purify Spot Punch Applications Blood, plant, insect, viral and bacterial analysis Genetic identification Ideal for clones Diagnostic and clinical applications Biosafety, food safety and environmental analysis HLA typing Animal breeding studies Molecular identification

FTA A Highly Flexible Technology used Widely in a Range of Industries FTA technology has been embraced by a wide range of industries across the globe. Pharmaceutical companies use FTA to collect and archive human DNA samples for clinical drug trials. Law enforcement agencies use FTA to collect and archive DNA samples from convicted offenders. Nature conservationists use FTA to collect bird DNA from jet engines to determine the flight patterns of specific species. Scientists hunting for new plant species use FTA in the field to collect and safely transport samples. Governmental agencies use FTA to sample food products while farmers use FTA to track diseases within multiple herd generations. While the range of applications is large, they all share a common element: simplicity. Whatman FTA helps scientists speed their research and achieve their goals. Use with virtually any cell type The following is a partial list of the cell types that can be applied to FTA Cards: Blood Cultured cells Buccal cells Plant tissue Bacteria Plasmids Micro-organisms Solid tissue Viral particles M13 plaques FTA Cards are available in either white or pink (Indicating) formats. White FTA Cards are recommended for blood samples, plant tissues and other easily identified samples. Indicating FTA Cards are pink and turn white upon sample addition. Indicating FTA Cards are recommended for buccal cells, cultured cells and other clear samples. Store nucleic acids at room temperature for years Genomic DNA stored on FTA Cards at room temperature for more than 14 years has been successfully amplified by PCR. No other product can make that claim. FTA Cards offer a compact room temperature storage system that reduces the need for precious freezer space, improves sample accessibility and reduces storage costs. Captured nucleic acid is ready for downstream applications in less than 30 minutes Captured nucleic acid is ready for purification when you are. Just take a sample disk from the FTA Card, wash with FTA Purification Reagent and rinse with TE -1 buffer. The washed disk is ready to use in applications such as PCR, RFLP analysis and RT-PCR. FTA DNA PURIFICATION PROTOCOL Sample Application Apply specimen and allow to dry completely. Disk Removal Punch a disk out of the sample area on the FTA Card. FTA Purification Reagent Washes Place the disk in a PCR tube and wash three times with FTA Purification Reagent. Discard used reagent after each wash. TE -1 Rinses Wash twice with TE -1 buffer (10 mm Tris, 0.1 mm EDTA, ph 8.0) and discard used buffer after each wash. Drying Step Dry disk in PCR tube. Direct to PCR Add PCR master mix directly to the disk and amplify.

FTA Cards, Reagent, Accessories and Kits FTA Cards FTA Cards are available in 1, 2, 3 and 4 part configurations. Custom configurations are available upon request. FTA Classic Card Four sample areas for storage of up to 500µL whole blood or 100µL plant homogenate per card. Convenient for multiple applications of the same specimen or collection of multiple animal or plant samples. Also available in Indicating (pink) FTA format. FTA Mini Card Two sample areas for storage of up to 250µL whole blood or 50µL plant homogenate per card. Convenient for protocols that require different locations for testing and archiving samples. Also available in Indicating (pink) FTA format. FTA Micro Card One sample area for storage of up to 125µL whole blood or 25µL plant homogenate per card. Recommended when only one sample is needed. Also available in Indicating (pink) FTA format. FTA Gene Card Three sample areas in a card frame for storage of up to 225µL whole blood or 30µL plant homogenate per card. Can be run in most automatic dispensing/pipetting systems when used with the FTA Gene Card Tray. CloneSaver Card Designed for the collection, storage and purification of plasmid and BAC DNA from bacterial clones. DNA is stable at room temperature for at least 5 years (real-time data). Available in a 96 well format for high throughput applications. PlantSaver FTA Card Plant friendly FTA Card, in a Classic Card format. Features a laminated flap that allows you to vigorously pound the plant sample into the FTA matrix without damaging the FTA Card. FTA Reagent, Accessories and Kits FTA Purification Reagent Removes heme, PCR inhibitors and other potential contaminants to ensure superior quality DNA for downstream analysis. FTA Gene Card Tray Holds two FTA Gene Cards for use in automatic liquid handling systems. FTA Kit Includes a 25-card supply of FTA Micro Cards; two vials of purification reagent (25mL); two Harris Uni-Core Punches; a cutting mat and instructions. FTA Starter Pack Provides a sample of FTA products, including one FTA Classic Card; one FTA Mini Card; one FTA Micro Card; one Indicating FTA Mini Card and one Indicating FTA Micro Card. Pack also includes two foam-tipped applicator swabs; one multi-barrier pouch and desiccant; one vial of purification reagent (25mL); two Harris Uni-Core Punches; a cutting mat and instructions. Sterile Foam Tipped Applicator Easy-to-use applicator for the non-invasive collection and transfer of buccal cells to FTA Cards. Harris Micro Punches, Disposable Uni-Core Punches and Cutting Mat For the precise sample disk removal from FTA Cards. The 1.2mm punches are recommended for use with whole blood and samples with high DNA content. The 2.0mm punches are recommended for use with buccal cells, plasmids and samples with lower DNA content. Multi-Barrier Pouches For transporting or storing FTA Cards. Protects cards from environmental contamination. Tamper-evident seal maintains sample security for forensics samples. A resealable pouch is also available when multiple access to FTA Cards is needed. FTA Card Mailer A rigid protective card mailer for transporting FTA Cards without biohazard labelling. Storage Desiccant Packets Ensure that FTA Cards remain dry during transport or storage. Contains indicator that changes colour to verify moisture absorption. CloneSaver Starter Kit Includes two CloneSaver Cards; two Harris Uni-Core Punches (2mm); a cutting mat and instructions.

FTA Mouse Tail DNA Whatman Raccolta, trasferimento, stoccaggio e purificazione del DNA da code di topo FTA della Whatman offre un valido e semplice metodo per la preparazione di campioni per analisi di DNA da code di topo. Occorre, infatti, semplicemente applicare un campione di sangue o di tessuto dalla coda del topo alla matrice FTA. Il DNA viene catturato e stabilizzato all istante, consentendone lo stoccaggio per un periodo indefinito a temperatura ambiente, da analizzare in qualsiasi momento. Trasporto sicuro e purificazione in 30 minuti rendono la tecnologia FTA un indispensabile strumento di ricerca. Caratteristiche Principali Metodo rapido e versatile di raccolta I campioni sono depositati direttamente sulla matrice FTA. Il DNA è purificato sulla FTA Card in tre steps semplici, in un singolo tubo, a temperatura ambiente. Non sono necessarie sostanze chimiche tossiche. Il DNA rimane fissato sulla matrice, pronto per la PCR. Adatto a qualsiasi metodo Le FTA Cards si adattano alle diverse tipologie di campione : preparazione di campioni di sangue o di campioni di tessuto di coda di topo previo pretrattamento mediante digestione enzimatica. E sufficiente applicare direttamente i campioni sulla matrice FTA e non è necessaria alcuna ulteriore purificazione né con sostanze tossiche (metodi quali fenolo/cloroformio) né con altri metodi che possono richiedere lunghe incubazioni. Stoccaggio a temperatura ambiente e trasferimento sicuri Gli acidi nucleici vengono automaticamente stabilizzati senza la necessità di congelarli. FTA inattiva gli agenti patogeni potenzialmente nocivi rendendo i campioni sicuri per la loro manipolazione in laboratorio. Inviare i campioni a colleghi o al laboratorio centrale è veramente facile con FTA Cards. Occorre solo inviarli per posta! Automatizzabile L utilizzo di sistemi automatici accelera le fasi di preparazione di diversi dischi prelevati contemporaneamente, ottimizza le risorse disponibili e standardizza la purificazione di DNA. I dischi prelevati dalla FTA Card possono essere sottoposti alle fasi di lavaggio e preparazione per la PCR su una diversi tipi di strumenti presenti in commercio (liquid handling robot). Tre semplici passaggi operativi per ottenere un DNA puro Dispensare il campione Purificare Prelevare un piccolo disco (punch) dal Campione Applicazioni Screening di topi transgenici Identificazione genetica Studi di allevamento Applicazioni diagnostiche PCR Analisi SNP Whole genome amplification

PROTOCOLLO APPLICATIVO DEL FTA MOUSE TAIL DNA Dispensare il campione Tagliare approssimativamente 1,5cm di coda del topo, stringere gentilmente perchè il sangue venga alla superficie del taglio, appoggiare leggermente la FTA Card sul sangue. Oppure applicare la raccolta direttamente sull FTA Card. Lasciare asciugare completamente. Prelevare un piccolo disco Prelevare (mediante perforazione) un piccolo disco dal campione depositato sulla FTA Card. Purificazione con Reagente di Purificazione FTA (Purification Reagent FTA) Mettere il disco di campione in un tubo per PCR ed effettuare tre lavaggi con il Reagente di Purificazione FTA. Eliminare la soluzione utilizzata dopo ciascun lavaggio. Lavaggio con soluzione tampone TE-1 Effettuare due lavaggi con la soluzione tampone TE-1 (10mM Tris, 0,1 mm EDTA, ph 8,0). Eliminare la soluzione usata dopo ciascun lavaggio. Essiccazione Lasciar asciugare il disco nel tubo PCR. PCR Aggiungere la master mix per la PCR direttamente al disco e iniziare la reazione di amplificazione. Qualità Whatman Whatman, azienda leader nelle tecnologie per la separazione, è nota nellà comunità scientifica per prodotti e soluzioni innovative contraddistinte da un elevata qualità. Il nostro istinto per la semplificazione accelera la velocità di fare nuove scoperte, riduce i costi e risparmia tempo. Inoltre, per accentuare ulteriormente l impegno nel soddisfare le specifiche esigenze dei nostri clienti, Whatman è organizzata in quattro aree distinte: LabScience, BioScience, MedTech Diagnostica e MedTech Dispoitivi. Ulteriori informazioni sono disponibili collegandovi al sito www.whatman.com. FTA, GenSpin e Whatman sono marchi registrati della Whatman Group. INFORMAZIONI PER L ORDINE DAL CATALOGO WHATMAN No. Cat. Descrizione Quantità per Confezione WB120061 FTA Starter Pack 1 WB120204 FTA Reagente di Purificazione 500mL WB120205 FTA Classic Card (non-indicatrice) 100 WB120055 FTA Mini Card (non-indicatrice) 100 WB120210 FTA Micro Card (non-indicatrice) 100 WB120208 FTA Gene Card 100 WB100007 Harris Micro Perforatore 2.0mm 1 WB100008 Harris Micro Perforatore 2.0mm Tip 1 WB100029 Harris Uni-Core 2.0mm Perforatore 4 WB100010 Busta Multi-Barrier (grande) 500 WB100011 Busta Multi-Barrier (piccola) 500 WB120216 FTA Gene Card/Busta/Essiccante 1000 WB120217 FTA Classic Card/Busta/Essiccante 1000 WB100026 2.0mm Plunger di Ricambio 1 WB100003 Essiccante (1g) 1000 WB100016 Busta Postale per FTA Card 50 WB100020 Tagliere di Ricambio 1 WB100030 Supporto per FTA Gene Card 20 WB120005 GenSpin Kit di Purificazione DNA 50 Purificazioni Purificazione del DNA in meno di 25 minuti! Il GenSpin Kit di Purificazione del DNA della Whatman rappresenta un metodo estremamente semplice per purificare il DNA a singolo filamento da campioni di sangue e cellule in coltura. Il campione è subito pronto per la PCR a partire anche da piccoli volumi del campione di partenza. North America Europe Japan Asia Pacific Whatman Inc 9 Bridewell Place, Clifton, NJ 07014 Technical Support: 1-800-922-0361 Customer Service: 1-800-631-7290 Outside US: 973-773-5800 Fax: 973-773-0168 E-mail: info@whatman.com Whatman International Ltd Springfield Mill James Whatman Way Maidstone Kent ME14 2LE, UK Tel: +44 (0)1622 676670 Fax: +44 (0)1622 691425 E-mail: information@whatman.com Whatman Japan KK Daiwa Ueno Building 1F 6-10 Ueno 5-chome, Taito-ku Tokyo 110-0005, Japan Tel: +81 (0)3 3832 6707 Fax: +81 (0)3 3832 6457 E-mail: japaninfo@whatman.com Whatman Asia Pacific Pte Ltd 171 Chin Swee Road #08-01 San Centre Singapore 169877 Tel: +65 6534 0138 Fax: +65 6534 2166 E-mail: wap@whatman.com GenSpin TM Kit di Purificazione DNA Whatman Leaders in Separations Technology www.whatman.com Cat No. S9036-783

FTA Plant DNA Whatman Raccolta, trasferimento e purificazione del DNA di Piante In laboratorio o in mezzo ad una foresta, FTA della Whatman offre un valido e semplice metodo per la preparazione di campioni per analisi di DNA di piante. Occorre, infatti, semplicemente applicare sulla matrice FTA il campione vegetale tal quale o dopo averne ottenuto un omogenato. Il DNA viene catturato e stabilizzato all istante, consentendone lo stoccaggio per un periodo indefinito a temperatura ambiente, da analizzare in qualsiasi momento. Trasporto sicuro e purificazione in 30 minuti rendono la tecnologia FTA un indispensabile strumento di ricerca. Caratteristiche Principali Raccolta semplice di campioni sul campo La stabilità degli acidi nucleici a temperatura ambiente e la protezione dalla degradazione rendono la tecnologia FTA uno strumento ideale per la raccolta di campioni sul campo. Meno campione E possible utilizzare anche solo le foglie più giovani, riducendo il tempo di crescita necessario e accelerando la ricerca. Stoccaggio a temperatura ambiente e trasferimento sicuri Consente di raccogliere ovunque il DNA dalle piante, di trasportarlo al laboratorio e purificarlo solo quando necessario. Purificazione rapida Il DNA viene purificato sulla FTA Card in tre semplici steps, in un singolo tubo, a temperatura ambiente. Non sono necessarie sostanze chimiche tossiche, inoltre rimanendo il DNA fissato sulla matrice è subito pronto per la PCR. Se il DNA è richiesto in soluzione, usare GenSpin Plant la Whatman. Ideale per automatizzazione L utilizzo di sistemi automatizzati accelera le fasi di manipolazione di multiple FTA Cards, riducendo i tempi di lavoro e standardizzando la purificazione del DNA. I dischi prelevati dalla FTA Card possono essere sottoposti alle fasi di lavaggio e preparazione per la PCR su diverse tipologie di strumenti presenti in commercio (liquid handling robot). Tre semplici passaggi operativi per ottenere puro DNA da piante Purificare Depositare Campione Perforare un Disco di Campione Applicazioni Analisi del DNA di piante mediante saggi PCR Selezione di colture mediante marker genetici Identificazione delle varietà vegetali Analisi di filogenesi Amplificazione di loci genetici LCN (low copy number loci) Invader assay Multiple displacement amplification Identificazione di specie transgeniche

PROTOCOLLO APPLICATIVO DEL FTA PLANT DNA Dispensare il campione Depositare il campione vegetale o l omogenato sulla FTA Card. Lasciarlo asciugare completamente. Soluzione tampone TE-1 Lavare due volte con soluzione tampone TE-1 (10mM Tris, 0,1 mm EDTA, ph 8,0). Eliminare la soluzione tampone usata dopo ciascun lavaggio. INFORMAZIONI PER L ORDINE DAL CATALOGO WHATMAN No. Cat. Prelevare un piccolo disco (punch) dal Campione Prelevare mediànte perforazione un disco dalla matrice FTA impregnata di materiale vegetale. Purificazione con Reagente di Purificazione FTA (Purification Reagent FTA) Mettere il disco di campione in un tubo per PCR ed effettuare tre lavaggi con il Reagente di Purificazione FTA. Eliminare la soluzione utilizzata dopo ciascun lavaggio. Descrizione Essiccazione Lasciar asciugare il disco nel tubo PCR PCR Aggiungere la master mix per la PCR direttamente al disco e iniziare la reazione di amplificazione. Quantità per Confezione WB120061 FTA Starter Pack 1 WB120204 FTA Reagente di Purificazione 500mL WB120205 FTA Classic Card (non-indicatrice) 100 WB120055 FTA Mini Card (non-indicatrice) 100 WB120210 FTA Micro Card (non-indicatrice) 100 WB120208 FTA Gene Card 100 WB100005 Harris Micro Perforatore 1.2mm 1 WB100006 Harris Micro Perforatore 1.2mm Tip 1 WB100028 Harris Uni-Core 1.25mm Perforatore 4 WB100010 Busta Multi-Barrier (grande) 500 WB100011 Busta Multi-Barrier (piccola) 500 WB120216 FTA Gene Card/Busta/Essiccante 1000 WB120217 FTA Classic Card/Busta/Essiccante 1000 WB100025 1.2mm Plunger di Ricambio 1 WB100003 Essiccante (1g) 1000 WB100016 Busta Postale FTA Card 50 WB100020 Tagliere di Ricambio 1 WB100030 Supporto per FTA Gene Card 20 WB120046 GenSpin Plant Kit 50 Purificazioni SWB120046 GenSpin Plant Kit di Prova 5 Purificatzioni Avete bisogno di DNA in soluzione? Questo kit di semplice utilizzo è ideale per la rapida preparazione di DNA a doppio filamento da piccole quantità di materiale vegetale, per analisi PCR. Il materiale raccolto dalle piante può essere omogeneizzato a temperatura ambiente e purificato in meno di 30 minuti. Qualità Whatman Whatman, azienda leader nelle tecnologie per la separazione, è nota nellà comunità scientifica per prodotti e soluzioni innovative contraddistinte da un elevata qualità. Il nostro istinto per la semplificazione accelera la velocità di fare nuove scoperte, riduce i costi e risparmia tempo. Inoltre, per accentuare ulteriormente l impegno nel soddisfare le specifiche esigenze dei nostri clienti, Whatman è organizzata in quattro aree distinte: LabScience, BioScience, MedTech Diagnostica e MedTech Dispoitivi. Ulteriori informazioni sono disponibili collegandovi al sito www.whatman.com. FTA, GenSpin e Whatman sono marchi registrati della Whatman Group. North America Europe Japan Asia Pacific Whatman Inc 9 Bridewell Place, Clifton, NJ 07014 Technical Support: 1-800-922-0361 Customer Service: 1-800-631-7290 Outside US: 973-773-5800 Fax: 973-773-0168 E-mail: info@whatman.com Whatman International Ltd Springfield Mill James Whatman Way Maidstone Kent ME14 2LE, UK Tel: +44 (0)1622 676670 Fax: +44 (0)1622 691425 E-mail: information@whatman.com Whatman Japan KK Daiwa Ueno Building 1F 6-10 Ueno 5-chome, Taito-ku Tokyo 110-0005, Japan Tel: +81 (0)3 3832 6707 Fax: +81 (0)3 3832 6457 E-mail: japaninfo@whatman.com Whatman Asia Pacific Pte Ltd 171 Chin Swee Road #08-01 San Centre Singapore 169877 Tel: +65 6534 0138 Fax: +65 6534 2166 E-mail: wap@whatman.com GenSpin Plant Kit di Purificazione DNA Whatman Leaders in Separations Technology www.whatman.com Cat No. S9036-784

FTA Author Title Year Publication Comment/Summary Keywords Aranda et al. FTA Technology, Unique Formats for 2001 Poster: Promega 12th Demonstrates FTA for downstream processes such as DNA elution, hair root, the Collection, Shipment, Archiving and Processing of Biological Samples International Symposium on Human ID from FTA, hair digestion, STR, extract DNA, microsatellites, human id, forensics multiplex PCR for human identification. Sample types buccal, human, blood, include hair roots, buccal cells, and blood restriction digest DNA Aranda et al. A Simple Device for the Efficient Transfer of Buccal Swab Cells onto FTA Paper 2003 Poster: Promega 14th International Symposium on Human ID, Phoenix AZ Describe how the SAMPACT device works. SAMPACT is a plastic cassett containing an FTA card that induces and equal pressure on a foam swab to get even transfer of buccal cells onto the FTA card forensic, databasing, human id Aranda et al. Alkaline Extraction of DNA from FTA Paper Spotted with Buccal Epithelial Cells and Whole Blood 2004 Poster: Promega 15th International Symposium on Human ID, Phoenix AZ Alkaline conditions were used to extract DNA from FTA DNA in solution, real time cards with blood or buccal cells applied to them. Extracted PCR, extracted DNA DNA was quantitated using the real time PCR kit Quantifiler from ABI. For blood samples extracted DNA from a 3mm punch ranged from 74 to 206 ng/punch; 7mm punches ranged from 165ng to 1.57ug. For buccal samples extracted DNA from a 3mm punch ranged from 35-52ng/punch; 7mm punches ranged from 31-109ng/punch. Babu Standing Orders - Institute of Aviation Medicine Aircrew DNA Repository 2002 Royal Australian Air Force Frame work as to how to administer and maintain aircrew's DNA repository for use in identification following an aircraft accident. How and when to retrieve archived samples. DNA repository, military, air force, disaster victim identification, blood samples, storage conditions, human id Baron et al. Parentage Testing and Effect of Misidentification on the Estimation of Breeding Value in Gir Cattle. 2002 Genetics and Molecular Biol 25(4):389-394. FTA used to collect blood samples from cattle. 2mm PCR, microsatellites, punches were washed and the DNA amplified by PCR in cattle, blood, parentage, 40 ml reactions directly. Also DNA from 6mm FTA punches genetic identity, breeding, were extracted using Chelex. dairy cows, Gir, genetic value of bulls, animal biotechnology Barton, JC, R Swada-Hirai, BE Rothenberg and RT Acton Two Novel Missense Mutations of the HFE Gene (I105T and G93R) and Identification of the S65C Mutation on Alabama Hemochromatosis Probands. 1999 Blood Cells, Molecules and Diseases 2 5(9)146-154. Note: a proband is a person with hemochromatosis a disease where there is too much iron in the blood, giving blood is a "treatment". Genomic DNA from saliva samples collected from volunteers on FTA for this study. Regions of the HFE gene were amplified by PCR then analyzed by sequencing and dot-blot hybridizations. human, population study, blood disease, saliva, buccal cell, FTA Reference DatabaseJun05 1

Beck et al. Beck and Frenkel Simple, Sensitive, and Specific Detection of Human Immunodeficiency Virus Type 1 Subtype B DNA in Dried Blood Samples for the Diagnosis in Infants in the Field Genotyping Kits for the Detection of HIV-1 pol Drug Resistance Mutations by an Oligonucleotide Ligation Assay 2001 Journal Clinical Microbiology 39: 29-33 2003 NIH AIDS Research and Reference Reagent Program Easy collection in the field for HIV testing in a central lab. Depicts High Sensitivity. Instructions for a kit to analyze HIV pol DNA. Whole blood collected on FTA (pg 14) can be used in the oligonucleotide ligation assay (OLA). These instructions also provide a good overview as to what the OLA is; briefly OLA is a way to identify a point mutation in a gene. A segment of the DNA is amplified by PCR then 2 oligonucleotides are hybridized to the PCR product. The 2 oligos are then ligated with a ligase enzyme. Only when a specific mutation is present will the oligos ligate and are detected. archiving, human, blood, PCR, HIV, sensitivity, specific, diagnosis, infection, pol, dried blood spots, diagnostics point mutation, PCR, HIV, pol gene, diagnostics Becker et al. Real-time PCR for detecting Trypanosoma brucei in human blood samples 2004 Diag. Microbiol. Inf. Dis. 50:193-199. Blood spiked with dilutions of cultured T. brucei and blood from patients with sleeping sickness were spotted to FTA cards. Punches of 2.0mm were washed according to the standard protocol then the DNA extracted using Chelex 100 resin. For real time PCR 4µl of the Chelex supernatant were included in the 10µl PCR mix. pathogens, disease diagnostics, human, bacteria, real-time PCR Belgrader & Automated Sample Processing Using Marino Robotics for Genetic Typing of STR Polymorphisms by Capillary Electrophoresis Belgrader et al. Automated DNA Purification and Amplification from Bloodstain Cards Using a Robotic Workstation Bever et al. Implementation of Laboratory Automation for the analysis of STR loci 1997 Lab. Robotics & Automation 9: 3-7 Use of FTA to rapidly produce template suitable for genotyping ID on an automated platform. 1995 BioTechniques 19: 427-432 Blood sample processing on FTA using an automated (Beckman) platform.. 1997 Poster 8th International Symposium on Human ID, The isolation of DNA using FTA & Rosys robotic workstation. robotic, automation, PCR, high-throughput, HT, blood, human, 96- well, human id, forensics, automation robotic, Biomek 1000, blood, human, automation human, blood, PowerPlex, DNA Plate, human id, forensics, automation Beyrer et al. Molecular Epidemiology of HIV-1 Among Northern Thai Drug Users: Implications for Vaccine Efficacy Trials. 2003 Poster 10th Conference on Retroviruses and Opportunistic Infections FTA used to collect blood samples from drug using volunteers in an HIV study. HIV, genetic diversity, infection, virus, diagnostics, population study FTA Reference DatabaseJun05 2

Bhattacharya et Use of Reverse Transcription and al. PCR to Discriminate Between Infectious and Non-Infectious Hepatitis A Virus. 2004 J Virological Meth. 116:181-187. Hepatitis A virus (an RNA virus), released into cell culture RT-PCR, RNA, virus, media from infected cells was applied to FTA cards. The environmental, food RNA was eluted from the cards in Tris-EDTA buffer pathogen, food safety, containing 2-mercaptoethanol then preciptated with isopropanol in the presence of carrier trna. RNA was also prepared using Trizol. Detection was more sensitive for RNA prepared on FTA than with Trizol. Bilyeu KD and Beuselinck PR Biondi et al. Birus et al. Genetic Divergence between North American Ancestral Soybean Lines and Introductions with Resistance to Soybean Cyst Nematode Revealed by Chloroplast Haplotype Forensic Sample Processing using a Robotic Workstation: Automated Paper-Based Spotting of Whole Blood convicted Offender Samples and High Throughput DNA Isolation for STR Analysis. How High Should Paternity Index Be for Reliable Identification of War Victims by DNA Typing? 2005 Journal of Heredity doi: 10.1093/hered/esi087 2003 Poster Promega 14th International Symposium on Human Identification Study was to examine genetic diversity in soybean genotypes present in USDA collection using SSRs in the chloroplast genome. Leaf tissue was pressed onto FTA cards or DNA from leaf tissue was prepared using Qiagen Plant mini kit or Promega Wizard Magnetic 96 Plant kit. 1 FTA punch or 10-100ng of DNA was included in each PCR amplification. MultiPROBE II Forensic workstation used to spot whole blood to FTA Gene Card in Gene Card tray for long term archiving. Shows how FTA and automation fit into forensic lab work flow. 2003 Croat Med J 44:322-326. FTA used to collect blood samples from relatives of missing persons for genomic DNA extraction. Soybean, germplasm collection, chloroplast DNA, Gene Card, blood, automaton, forensic, human ID human, missing persons, ID, STR, PCR, skeleton, genetic match, chelex, genotyping, databasing, human id, forensics Blair et al. Evidence of Rickettsial and Leptospira Infections in Andean Northern Peru 2004 Am. J. Trop. Med. Hyg. 70(4): 357-363. Blood collected from study participants in vacutainer tubes with some being spotted onto FTA for genomic DNA analysis of bacteria from the genus Rickettsia. A nested PCR method was used to identify a Rickettsia specific gene, htra. field collection, human, blood, bacterial infections, environmental, diagnostic, population study Borys et al. Borys. S PCR Volume Reduction Study Using Bloodstained FTA Collection Cards and Capillary Electrophoresis Evaluation of High Throughput STR Technologies for Potential Implementation in the National DNA Databank 1998 Poster Promega 9th Annual Int Symp Of Human ID. 1999 MSc. Thesis Ottowa-Carleton Institute of Biology, Carleton University, Ottowa Canada Final PCR volume of 5µl gives good, reproducible, STR profiling results; cycles reduced form 25 to 23. Use of FTA to collect blood, buccal cells for DNA analysis and databasing. Reducing PCR from 25 ml to 5 ml increases sensitivity 9-fold. blood, buccal, human, RCMP, STR, PCR, forensics, human id DNA bank, repositories, database, DNA typing, genotyping, human, blood, buccal, databank, forensics, human id FTA Reference DatabaseJun05 3

Bosman et al. Reverse Line Blot: A diagnostic tool to detect blood parasites. 2002 Presentation: 31 st Annual Conference of the Parasitological Society of Southern Africa (abstract # Blood from various animal species was applied to FTA or collected in vacutainers with various anti-coagulants. DNA was extracted or FTA punches prepared with the standard procedure and analyzed by PCR for the presence of Babesia or Theileria parasites. Parasites, whole blood, animal, PCR, animal biotechnology 17) Both et al. FTA Paper, DNA, Time, and the Profiler 2000 Web Article from Forensic Science Center Adelaide, Australia Review of FTA for the collection & archiving of offender blood samples.. RT storage for 9 years w/out signal loss. Punches from blood stained FTA could be PCR'd without washing, but punches containing buccal DNA typing, blood, buccal, human, archive, PCR, STR, integrity, human id, forensics cells required washing for successful PCR. ABI ProfilerPlus PCR kit was used in this study. Budowle et al. DNA Typing Protocols: Molecular Biology and Forensic Analysis. 2000 BioTechniques Books Publication, Eaton Publishing, Natick MA Description of FTA as a sample collection medium for blood and buccal cells. Protocol for preparing FTA punch for RFLP or PCR-based analysis. Shows photos of CEP swab aka OmniSwab and Gene Guard Swab aka genomic DNA, forensic, DNA typing, PCR, CEP swab, human id, forensics Foam tipped applicator for collecting buccal cells. Burgoyne Convenient DNA Collection and Processing: Disposable Toothbrushes and FTA Paper as a Nonthreatening Buccal-Cell Collection Kit 1997 Poster Promega 8th International Symposium on Human ID Use of a cytobrush to collect buccal cells to deposit on FTA buccal cell, human, buccalbrush, food coloring, forensics, human id Compatible with Automatable DNA Burgoyne & Hallsworth Processing Studies with FTA 1995 Presentation 5th International Symposium on Human ID FTA kills HSV within 5 sec. FTA prevents fungal growth on blood spots held at body temp and high humidity. FTA can be used to detect HSV, CMV (DNA viruses) and Hepatitis C (RNA virus) and protect samples from accelerated and natural aging. HSV, Herpes Simplex Virus, 903, 3MM, FTA923, Cytomegalovirus, CMV, Hepatitis C, DNA virus, RNA virus, diagnostics Castilho et al. DNA Template Preparation and Archiving for Blood Group Genotyping of Remotely Located Patient Populations 2001 Poster: American Association of Blood Banks, San Antonio 2001 FTA cards were used for blood group genotyping using whole blood, plasma, and amniotic fluid. The cards were used for collection in remote areas and shipped back to central lab for testing. storage, transportation, human, PCR, RFLP, population study, human id, diagnostics FTA Reference DatabaseJun05 4

Castilho et al. Genotyping for DO A/DO B Facilitates Transfusion of Sickle Cell Disease Patients 2001 Poster: American Association of Blood Banks, San Antonio 2001 Blood samples from patients who received a transfusion was spotted to FTA, shipped back to central lab for PCR analysis. Dombrock gene, RFLP, human, blood, genotyping, population study, human id, diagnostics Cerda-Flores et al. Maximum Likelihood Estimates of Admixture in Northeastern Mexico Using 13 Short Tandem Repeat Loci. 2002 Am J Hum Biol 14:429-439. Blood from people in Northeastern Mexico were collected and FTA and analyzed by PCR for 13 STRs. Data was analyzed to determine lineage contributions from Europe, American Indian and African origin. PCR, 1 mm punch, STR, human, blood, population study, human id Chappuis et al. Chu et al. Options for Field Diagnosis of Human African Trypanosomiasis Survey of Nepalese Animals for the Presence of Cyclospora Cayetanensis 2005 Clin. Microbiol. Rev. 18(1)133-146. 2003 Poster: Am Assoc. Microbiology, Washington DC FTA is described for collecting blood, lymph node fluid or CSF for detection of trypanosome DNA. FTA is preferred over untreated filter paper because FTA protects DNA from degradation. FTA used to collect animal fecal samples to detect protozoan parasites protozoa, protozoan, microorganism, disease diagnostics, PCR, disease diagnostics, fecal samples, animals, detection, PCR, nested PCR Chu et al. Detection of Cyclospora Cayetanensis in animal fecal isolates from Nepal using and FTA filterbase polymerase chain reaction method 2004 Am. J. Trop. Med. Hyg 71(4):373-379 FTA was used to collect fecal samples of dogs,chickens and monkeys to detect protozoan parasites capable of being transmitted to humans and causing diarrheal illnesses. Fecal samples were collected in 2.5% potassium dichromate (to stabilize the protozoan oocytes). Stool specimens 10-20ul were spotted disease diagnostics, fecal samples, animals, detection, PCR, nested PCR directly onto FTA and dried on a hot block at 56 o C. The disks were washed as usual then amplified. Connolly et al. Automated methods for processing samples stored on FTA paper 1997 Poster 11th International Symposium on Human ID, Biloxi, LA FTA has proven itself to be a robust & versatile method for collection, storage & analysis of DNA. With (semi-) automated methods is capable of high- throughput analysis. plate washers, throughput, automation Connolly et al. D12 archiving & recovery of bacterial plasmids using FTA paper 1999 Poster 13th International Mouse genome Conference Archiving & recovery of clones ranging from 2Kb to 227Kb. plasmid DNA, freezer, storage, CloneSaver, 96- well, M13 sequencing, genomics FTA Reference DatabaseJun05 5

Crabbe A novel method for the transport and analysis of genetic material from polyps and zooxanthellae of scleractinian corals. 2003 J. Biochem. Biophys. Methods 57: 171-176 Both the polyps (the soft living part of coral, about the size of a pencil eraser) and the symbiotic algae (zooxanthellae) that are part of coral colonies were collected and put on FTA. Fragments were ground and applied to FTA. DNA was extracted using Qiagen DNeasy. Ribosomal genes were PCR amplified for species identification. FTA excellent for field collection and transport of samples from tropical locations. coral, algae, identification, ribosomal genes, extracted DNA, tropical, field collection, room temperature transport, plant Davis et al. DNA Tests in Prolific Sheep from Eight Countries Provide New Evidence on Origin of the Booroola (FecB) Mutation. 2002 Biology of Reproduction 66:1869-1874. Blood samples from sheep collected onto FTA for DNA preparation. FTA disks (1.2mm) were amplified by PCR and the amplicons subjected to RFLP analysis. PCR amplicons prepared from FTA disks containing sheep DNA were also subjected to TaqMan Assays. animal biotechnology, blood, mutation analysis Davis, et al. A Novel Genvault Multiwell Plate Format for Whatman FTA in Robotic, High Throughput DNA Archiving 2003 Poster: Society for BioMolecular Screening Meeting Describes the Genvault system and the EasyClone 384 well plate EasyClone 384 well Plate Del Rio Cost Effectiveness in Sample Processing using the FTA Treated Stain Card for High Throughput 2001 Poster from Promega 8 th International Symposium on Human Identification Important for showing the cost-effectiveness of FTA for blood collection and processing compared to vacutainer collection. offender specimens, archive, collection, DNA database, transport, sample tracking, human, blood, buccal, human id, forensics Del Rio et al. Del Rio, S. Reusing the Same Blood-Stained Punch for Sequential DNA Amplifications and Typing Amplification of DNA from Bone Marrow Aspirate and Cervical Smears on FTA Blood Stain Cards 1996 BioTechniques 20: 970-974 Demonstrates that not all NA bound to FTA cannot be removed with heat, and punches can be re-used for many different PCR amplifications multiple times (4 sequential runs) 1997 Application Note from Fitzco Different sample types than blood processed successfully on FTA (either spotted or brushed on the cards) human, genotyping, databanks, human id, diagnostics, forensics, research diagnostic tests, diagnostics, Del Rio- LaFreniere et al New Method for Collection and Detection of K-ras Point Mutations from Tissue Specimens for Clinical Diagnostic Applications 1998 Poster: AMP Meeting Washington DC Samples of pancreatic tissue, ductal brushings, cells, cancer research, tissue fine needle aspirates or fluid were applied to FTA. FTA screening for mutation, cards were pressed to tissue samples. A 3mm punch diagnostics was taken and washed as normal. PCR master mix was added to the washed punch to detect mutation in the K- ras gene. Bile and fine needle aspirates showed the weakest signal on FTA but the others gave good PCR bands on gels. FTA Reference DatabaseJun05 6

Del Rio- LaFreniere SA, McGlennen RC A Unique Method of Detection of the Prothrombin 20210A (FactorII) Mutation Using the Simultaneous Allele Specific Amplification (SASA) 1998 Poster: 13th San Diego Conference on DNA Technologies in Human Detection Human Blood sample were collected on FTA and washed 3mm punches subjected to a specific PCR amplification procedure for detecting a mutation in the prothrombin gene. disease diagnostics, PCR, mutation research, Devost & Choy Mutation Analysis of Gaucher Disease Using Dot-Blood Samples on FTA Filter Paper 2000 American Journal of Medical Genetics 94: 417-420 Polymorphism detection of alleles implicated in Gaucher Disease on population bloods collected & stored on FTA. diagnostic tests, RFLP, mutation, genotyping, nested PCR, human, diagnostics, population study Dobbs et al. Dutton and Tieber Drescher & Graner Use of FTA Gene Guard Filter Paper for the Storage and Transportation of Tumor Cells for Molecular Testing A Modified Protocol for Sex Identification of In Ovo Avian Embryos and its Application as a Management Tool for Endangered Species Conservation Programs. PCR-genotyping of barley seedlings using DNA samples from tissue prints 2002 Archives Pathology and Laboratory Medicine 126: 56-63 Shows FTA for tumor cell collection, room temperature storage, transport, and DNA testing. Compares FTA with a Qiagen method. 2001 J Zoo Wildlife Med 32(2)176-Small amounts of blood were collected by syringe from blood vessels in developing bird eggs. The blood/fluid was 180. spotted to FTA and analyzed to determine the gender of the embryo. 2001 Plant Breeding 121, 228-231 Method to sample barley leaves for PCR analysis using FTA. Compares FTA to CTAB standard method; achieve comparable results quicker and easier. pathology, cancer, lymph node, cell suspensions, lung, breast, endometrium, ovary, kidney, thyroid, research, DNA banking blood, animal biotechnology, birds, chicken, pigeon, owl marker assisted breeding, plant, screening, leaves, high throughput, genotyping, field collection Dyer et al. Detection of Bacilli Spores Using PCR and FTA Filters 2003 Poster 103rd General Meeting American Society for Microbiology FTA used to isolate DNA from Bacilli spores for nested PCR detection. spores, Bacillus, food, clinical, environmental, Elliot Isolation of DNA Using FTA Blood Stain Collection Cards 1998 Poster 7th Annual International Symposium of Human ID FTA for forensic collection, archive, rapid purification tool. 9 months RT archiving. Did not let card dry after blood application, put it wet into pouch with desiccant without effecting PCR. RCMP, rapid collection, rapid extraction, human, saliva, buccal, STR, blood, storage conditions, forensics, human id FTA Reference DatabaseJun05 7

Elliot et al. Extraction of DNA from FTA Blood Stain Collection Cards for Construction of a Large STR National DNA Data Base 1997 Poster 8th International Symposium on Human ID, FTA tested for DNA stability, yield (peak height on a gel) ease of use and consistency with blood fresh, frozen, saliva and semen. Did not have good luck with purified DNA on FTA with standard wash protocol. RCMP, rapid collection, rapid extraction, human, saliva, buccal, STR, blood, storage conditions, forensics, human id Expert et al. Evaluation of Two Techniques for Extraction and Conservation of DNA of Various Quarantine Bacteria 2004 Poster EPPO Conference on Quality of Diagnosis and New Diagnostic Methods for Plant Pests. Noordwijkerhout, NL Extraction of DNA from plant disease bacteria by FTA compared to Q Biogene kit. Both methods able to detect gram (-) bacteria at low levels seen in plants. Both methods could only detect gram (+) bacteria at higher concentrations. plant disease, microbes, Fici et al. Sequence Based Typing of Blood Spots on Filter Paper After Extended Storage Forrest et al. Polymorphism at the Ovine b3- Adrenergic Receptor Locus: Associations with Birth Weight, Growth Rate, Carcass Composition and Cold Survival. Forrest et al. Two Rare Polymorphisms in the D8S1179 and D13S317Markers and Method to Mitigate Their Impact on Human Identification 1998 Poster Human Immunology Blood and lymphocyte samples spotted to FTA cards 59(1) 141 then processed for HLA & blood typing after long-term storage. PCR from FTA paper more robust than PCR from untreated filter paper. A treatment with Proteinase K (5 ml 10 mg/ml prok added to 495 ml FTA purification reagent containing punch; incubate 1 hr @ 60 o C)was required to amplify one class of the HLA. 2003 Animal Genetics 34:19-25. FTA used to collect blood from lambs. DNA on FTA punches were examined via PCR for differences in the sequence in the b3-adrenergic receptor gene which were then compared to phenotypic traits of the sheep. 2004 Croat Med J 45:457-460 Blood collected onto FTA and used in human genotyping with the commercially available kits. The researchers saw a new peak in two of the loci, sequenced the DNA and found DNA base changes that affected the STR profile. They make suggestions as to what that means for identifying humans in forensic cases. untreated filters, spin basket, 30 months, 2.5 years storage, Proteinase K, diagnostics, population study PCR-SSCP, polymorphisms, body weight, marker assisted breeding, animal ID, animal biotechnology human id, forensics, DNA polymorphisms, DNA sequencing, STR Fox et al. RT-PCR from Eukaryotic Cells Stored on FTA Archival Paper 2000 Poster Abstract: Plant and Animal Genome VIII San Diego Demonstrates FTA for collection, storage, processing of mrna for RT-PCR- mrna from human cells spotted onto FTA was stable for 1-2 mos room temp or > 10 mos at - 20 o C or -70 o C. RNA extraction, plant, BHK21 cells, HeLa cells, high copy number mrna, low copy number, FTA Reference DatabaseJun05 8

Franchina et al. Fujiwara et al. Fyfe et al. Polymorphism of the CD30 Promoter Microsatellite Repressive Element is Associated with Development of Primary Cutaneous Lymphoproliferative Disorders. Plasma DNA Microsattelites as Tumor-specific Markers and Indicators of Tumor Progression in Melanoma Patients. Assessment of trypanosome prevalence in FITCA high risk areas. 2005 Cancer Epidemiol Biomarkers Prev 14(5) 1322-1325. Blood from patients was collected on FTA for PCR amplification of sections of the CD30 gene. 1999 Cancer Res. 59:1567-1571. Blood collected from 76 cancer patients and 20 healthy donor volunteers and spotted to FTA for genomic DNA extractions and long term storage. 2003 ICPTV Newsletter 8 pg 25-26 September 2003 FITCA is an EU funded project to control tsetse infestation in Uganda. Blood from cattle (300-400) ear veins was collected directly onto FTA cards and transported back to the lab for analysis by PCR for 3 species of Trypanosome. Detection of parasite by PCR is significantly more sensitive than detection by microscopy. population study, cancer research cancer research, tumor progression, Loss of heterozygosity, human, diagnostic cattle, DNA, PCR, parasite, animal biotechnology, environmental Gaytmenn et al. Determination of the sensitivity and specificity of sibship calculations using AmpFI STR Profiler Plus. 2002 Int J Legal Med 116:161-164 Buccal cell samples were collected on FTA and used as template for the 9 STR Profiler Plus loci. These data were used to determine the likelyhood ratios of paternity and sibling relationships. Buccal cells, human, STR, paternity, sibling, population study, human id, forensics Goldsborough et al. Room Temperature Archiving of Plasmid Clones in an Automatable 96- Well Format 2000 Poster Abstract: Plant and Animal Genome VIII San Diego, USA Demonstrates use for BAC and plasmid clones. glycerol stocks, overnight cultures, resuspended colonies, purified DNA, 2 Kb, 200 Kb, CloneSaver, genomics Gutierrez- Corchero et al. Using FTA cards to store avian blood samples for genetic studies. Their application in sex determination 2002 Molecular Ecology Notes 2: 75-77 FTA cards were used to collect, store, and analyze blood from nestling and adult birds (Great Grey Shrikes) for sex determination. birds, DNA banks, wildlife ecology, PCR, population studies, blood, nucleated red blood cells, animal biotechnology Halbert et al Conservation Genetic Analysis of the Texas State Bison Herd 2004 J Mammalology 85(5):924-931 Blood samples were collected on FTA. Single 1.2mm punches were used in multiplex PCR amplifications in 5ul. animal ID, population study Hanes et al. Inactivation of Cryptosporidium parvum Oocytes in Fresh Apple Cider by UV Irradiation 2002 Appl Envir Microbiol. 68(8) 4168-4172 Apple cider containing parasitic oocytes was spotted on FTA and analyzed by PCR. gastroenteritis, fecal pellets, tissue, parasites, infection, environmental, food safety FTA Reference DatabaseJun05 9

Hansen and Blakesley Simple Archiving of Bacterial and Plasmid DNAs for Future Use 1998 Focus 20(3) 72-74 Bacterial genomic DNA was stored on an FTA card with successful PCR amplifications. Also, plasmid DNA was stored and transformed on FTA. Agrobacter tumefaciens, plant bacteria, E. cole, gram negative, gram positive Streptomyces, colonies, clones, genomics Hartman and Lampel Long Term Room Temperature Storage and Detection of Norovirus on FTA Filter Paper. 2004 Poster: 104th General Meeting of the American Society of Microbiology, New Orleans LA. Norwalk-like virus is a food borne pathogen. Norovirus is a Virus, RT-PCR, food leading cause of non-bacterial gasteroenteritis. Diagnosis borne pathogen, is almost exlusively done by detection of virus in stool detection, stool, feces, samples. Norovirus positive stool was diluted 1:10 with vomit, rapid analysis PBS and 200ul applied to FTA cards and dried overnight. Cards were stored in plastic Whirl-Pak bags for 6 and 12 months at room temperature. One sample was stored for 2 years. A 6mm disk of FTA (approx 6-7ul stool) was washed with 140ul sterile dist H 2 O. Viral RNA extracted with QiaAmp Viral RNA Mini Kit (Qiagen). All samples analyzed from FTA showed positive signal for Norovirus. FTA makes sample collection, storage and transportation easy and can be used to quickly detect Norovirus outbreaks. FTA can facilitate epidemiological investigations. Harvey, ML An alternative for the Extraction and Storage of DNA from Insects in Forensic Entomology. Henderson et al. The long term outcome of limbal allografts: the search for surviving cells. 2005 J Forensic Sci. 50(3) www.astm.org 2001 Br J Ophthalmol 85:604-609 Samples of various life stages of flies were applied to FTA. A 3 fragments of 320, 650 and 1270 bp of a gene encoded in the mitochondrial DNA were amplified by PCR to show the quality of the DNA. Authors found the 1.2mm punch to be optimal for PCR amplification; 2.0mm punch provided FTA disks were pressed against the corneas of eyes receiving limbal allografts to collect cell samples. The DNA from the cells were typed using 4 human STR markers to determine if they derived from the donor or the recepient. insects, forensics, entomology, mtdna impression cytology, human identification, ID, clinical, tissue grafts, diagnostic Hernandez et Comparative Molecular Study of al. Populations of Common Crossbill (Loxia curvirostra ) on the Iberian Peninsula and the Baleric Islands. Hernandez et al. Identification of Lanius Species and Subspecies using Tandem Repeats in the Mitochondrial DNA Control Region. 2003 Workshop WS02-P3 4th Conference of the European Ornithologists' Union Blood samples from birds collected onto FTA cards and prepared for PCR amplification of microsatellite markers. 2004 Ibis 146:227-230. Mitochondiral DNA analyzed from bird blood spotted to FTA microsatellites, avian, blood, animal biotechnology birds, mitochondiral DNA FTA Reference DatabaseJun05 10

Hide et al. A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper. 2003 BMC Ecology 3:7 A specific protozoan, Bleparisma japonicum, detected in environmental samples applied to FTA cards. Single cell detection levels achieved. Larger number of cells 2.5 and 5 cell equivalents on FTA inhibited PCR. Canal, river and pond water samples tested and B japonicum specifically detected in the presence of other protozoans. Protozoa, water samples, environmental samples, collection, storage Hickford et al. Diversity of the Ovine DQA2 Gene 2004 J. Anim Sci. 82:1553-1563. Blood from 2,000 sheep collected onto FTA cards. DQA2 gene was amplified by PCR cloned and sequenced. genetic analysis, agriculture, animal, Higgins et al. Sensitive and Rapid Identification of Biological Threat Agents 1999 Ann NY Acad Sci 894: 130-148 FTA was senstive for detecting Bacillis anthracis in buffer and whole blood samples anthrax, blood, biological threat agents Higgins et al. Detection of Francisella tularensis in Infected Mammals and Vectors Using A Probe Based Polymerase Chain Reaction 2000 Am J of Tropical Medicine and Hygiene 62:310-318 FTA used to collect microorganism in liver tissue smears, then detection carried out by TaqMan PCR. real-time PCR, mouse tissue, tick extract, insects, gram negative, infection, rapid detection, bioterrorism, purified DNA, blood, tail blood, Dermacentor reticulatus, Ixodes rincinus, Haemaphysalis concinna, PCR-EIA, nested PCR, environmental, parasite, animal biotechnology Ho et al. Outbreak of Cyclosporaiasis Associated with Imported Raspberries, Philadelphia, Pennsylvania, 2000. 2002 Emerg. Infect. Dis. 8(8) serial online FTA used to collect food samples to detect parasite infection. food safety, DNA extraction, ethyl acetate extraction, FDA, CDC, parasite oocytes, produce, environmental Howard et al. Standardizing Yields from Blood Card Punches Extracted with DNA IQ and the Biomek Liquid Handler 2004 Poster: 15th Promega Human ID Symposium, Phoenix AZ Punches of 3.2mm from BSD Duet from both FTA and 903 cards with blood samples were extracted with the Promega DNA IQ extraction system. Both yielded the same amount of blood but yields from FTA was linear as more blood punches were extracted. FTA yielded more DNA than 903 paper. Authors say storage conditions and DNA degradation must be taken into effect when creating a standard extraction method. human id, forensics, purified DNA, extracted DNA, FTA Reference DatabaseJun05 11

Hsiao et al. Application of FTA Sample Collection and DNA Purification System on the Determination of CTG Trinucleotide Repeat Size by PCR-Based Southern Blotting 1999 J. Clin Lab. Anal. 13: 188-193 Collection and processing tool for population samples for genotyping. FTA ability to preserve DNA against degradation is critical to this assay. gene mutation, myotonic dystrophy (DM), deazadgtp, long term storage, human, blood, lymphoblast cells, Hunter et al. The P28T Mutation in the GALK1 Gene Accounts for Galactokinase Deficiency in Roma (Gypsy) Patients Across Europe. 2002 Pediatr Res 51:602-606 Used FTA to collect blood from patients to do PCR on genomic DNA. gene mutation, galactokinase gene 1, human, blood, PCR, screening, microsatellites, Ibrahim et al. Real Time Microchip PCR for Detecting Single-Base Differences in Viral and Human DNA 1998 Analytical Chemistry 70: 2013-2017 TaqMan PCR demonstrated for samples presented to and stored on FTA cards infectious disease, genetic disorders, single nucleotide polymorphism, SNP, orthopoxvirus, cowpox, monkeypox, camelpox, vaccinia subspecies buffalopox, human, blood, Igoe et al. A Novel Automatable System for the Room Temperature Archiving and Recovery of DNA Clones 2001 Poster: 11th Genome Sequencing and Analysis Conference, San Diego, USA Demonstrates the capabilities of the CloneSaver card for storage of plasmid and BAC clones including phage inactivation, glycerol stock application, no crosscontamination, and archiving capabilities. phage inactivation, cross contamination, 3 yr plasmid archiving, Igoe et al. A High-Throughput System for Room Temperature Archiving and Processing of Plasmid and BAC Clones. 2002 Poster: 12th Genome Sequencing and Analysis Conference, Boston MA Demonstrates a prototype of a "hard" CloneSaver, automated sample application of plasmid DNA using the Biomek 2000 and the use of FTA as a template for Templiphi template amplification kit. HardCard, Biomek 2000, automated punch washing, rolling circle amplification, Igoe et al. Purification and Analysis of DNA from Solid-Phase Samples Using an Automated Platform. 2003 Poster: IX Plant and Animal Genome Conference, San Diego, USA FTA and CloneSaver punches were washed using the Biomek 2000 liquid handler in an automated fashion. Studies proved no cross contamination between wells. automated punch washing, plant DNA, blood, plasmid, transformation Igoe and Robbins Amplifying Stored Plasmids for Sequencing 2004 Genetic Engineering News 24(2): 34 Describes using Amersham's TempliPhi kit to amplify plasmid DNA stored on CloneSaver. Plasmid DNA was eluted from CloneSaver and amplified with TempliPhi. The amplified plasmid was cut with restriction enzymes or was sequenced using Applied Biosystems BigDye v 3.0 kit. Plasmid, clones, genomics, rolling circle amplification, FTA Reference DatabaseJun05 12

Igoe et al. Comparison of DNA Stability on Treated and Untreated Papers. Ivanov et al. The Experience of using the FTA Cards for Blood Samples Storage at Conducting the Molecular-and- Genetic Identification of Nonidentified Killed Persons - Victims of Military Actions in the Territory of the Chechen Republic Jankowski et al. The Comparison and Validation of FTA, CHELEX, and Qiagen Extraction Methods on Blood Spotted FTA Cards for the Analysis of the 13 Core STR Loci Johnson A PCR-Based Method for Detection of Shigella in Produce Washes. Jones et al. The Use of FTA for the Identification of Gram Positive and Spore-Forming Bacteria. Jou et al. Delineation of CTG Repeats and Clinical Features in a Taiwanese Mytotonic Dystrophy Family. 2004 Poster: American Academy of Purified DNA applied to FTA was protected from UV Forensic Science Dallas TX 2002 Sudebno-Meditinskaya Ekspertiza 45:20-27. 2000 Poster 11th International Symposium on Human ID, Biloxi, LA 2003 Presentation AOAC International Midwest Section Meeting and Exposition 2003 Poster: San Diego Conference: Cool Tools, Baltimore MD Oct 30 - Nov 1, 2003 2001 Proc. Natl. Sci. Counc. ROC(B) 25 (1) 40-44. damage compared to DNA on untreated filter paper. DNA was more stable on FTA over time and storage conditions compared to untreated filter paper. Blood applied to glass fiber media treated with FTA was stable after 5 mos where as DNA on untreated media was completely degraded and not amplifiable by PCR. PCR amplification of mitochondrial DNA; HVS-1 and -2 (hypervariable segments) of the D-loop. Analysis of PCR fragments from Profiler Plus system on ABI Prism 377. Describes benefit of room temperature storage over traditional storage methods. Blood on FTA stored from several weeks to 3.5 years. Describes benefit of FTA over phenol-detergent or Chelex extractions of profiling. untreated filters, DNA stability mtdna, missing persons, military, 1100 blood samples, human, DNA. Analysis of several methods of Processing FTA for STR extract DNA from FTA, Compare FTA to Qiagen Dnease Plant Mini Kit in preparing Shigella for PCR analysis from plant washes. Also compare BAX real-time PCR detection system to PCR and gel electrophoresis. Both FTA and Qiagen prepared a good template. Spore-forming bacteria and gram positive bacteria applied to FTA and analyzed by PCR for 16S RNA gene, esp and gdh genes. Bacterial genomic DNA could be detected at a dilution of 10-7 showing that FTA provides a sensitive level of detection. FTA used to collect patient buccal cells and prepare Real-time PCR, BAX automation detection system, bacteria, produce, pathogens, gram positive bacteria, spores, DNA for PCR amplification to detect genetic alterations buccal, human, in the myotonin protein kinase gene in myotonic dystrophy. clinical, gene mutation, FTA Reference DatabaseJun05 13

Karle et al. Novel filter-based technology for simple collection and preparation of PCR-ready plant DNA:applications for transgenic plant detection 2001 Poster: 11th Genome Sequencing and Analysis Conference Demonstrates processing plant samples for DNA analysis using FTA and GenSpin Kit for Plants alfalfa, Arabidopsis, Brassica, corn, cotton, cucumber, potato, diploid potato, rice, ryegrass, soybean, spinach, tomato, wheat, SSR amplification, transgenic, Bt-Cry3A, Rca gene, rbcl gene, SATT309, nematode resistance, GMO Karle et al. Application of FTA-based Technology for Simple Collection, Transport, Purification and Storage of PCRready plant DNA. 2002 Poster: Xth Plant and Animal Genome Conference, San Diego, USA Demonstrates the utility of FTA for collection, storage, simple DNA purification, and DNA identification of plants alfalfa, Arabidopsis, Brassica, corn, cotton, cucumber, potato, diploid potato, rice, ryegrass, soybean, spinach, tomato, wheat, SSR amplification, transgenic, Bt-Cry3A, Rca gene, rbcl gene, SATT309, nematode resistance, GMO Kerlin et al. Survival Advantage Associated with Heterozygous Factor V Leiden Mutation in Patients with Severe Sepsis and in Mouse Endotoxemia 2003 Blood 102(9)3085-3092 Whole blood collected from 1605 patients sent to Lilly Research Labs to analyze mutations in Factor V gene by PCR septic shock, mutation analysis, clinical trial Kline et al. Polymerase Chain Reaction Amplification of DNA from Aged Blood Stains: Quantitative Evaluation of the "Suitability for Purpose" of Four Filter Papers as Archival Media 2002 Analytical Chemistry 74: 1863-1869 NIST evaluated four different card matrixes for STR multiplex performance from dried blood stored on the cards for 19 months. Publication also demonstrates the ability to elute DNA off of FTA Cards using a Chelex extraction procedure. Blood stain paper, S&S 903, IsoCode, FTA, chelex, blood, human, FTA Reference DatabaseJun05 14

Kozlowski et al. SNP and Transgene Analysis Directly from Sample Collection Paper using the Invader DNA Assay. 2003 Poster: Plant & Animal Genomes XI Conference, San Diego, CA FTA used in Third Wave Invader Assay to detect SNPs in pigs/sheep and also transgenes in soybean/corn. Each reaction was a biplex, measuring 2 targets per reaction. S&S 903 and untreated blotting paper were also tested. Samples were blood or leaf material and 2mm punches were assayed in 96 well plates. Leaf punches were washed in 85% ethanol; blood punches were washed in water; then punches were incubated in water for 10 min at 95 o C. The Invader reagents were added and incubated 65 o C for 2-4hrs. Results from FTA cards were similar to results from genomic DNA purified via Gentra DNA kit; 903 and untreated blotting paper did not work well and require more optimization. Blood samples and soybean samples on FTA worked great, corn needs more optimization. SNP, porcine, ovine, blood, scrapie, stress syndrome, transgenic plants, GMO Krenke et al. Validation of a 10-Locus Flourescent Multiplex System 2002 J Forensic Sci 47(4):773-785 An STR kit was validated in several forensic labs under a variety of conditions. Dried blood on FTA either amplified directly or was extracted by Promega DNA IQ system. FTA punches performed better if the standard number of PCR cycles was 10/20 to 10/22. forensic, validation experiments, human ID Kuboki et al. Loop-Mediated Isothermal Amplification for Detection of African Trypanosomes 2003 J. Clin Microbiol 41(12)5517-5524. FTA used to collect blood from mice and cattle infected with protozoan parasites. Washed FTA punches were used in both PCR and a non-pcr based amplification reaction, an isothermal (all at one temperature) reaction parasites, diagnostic, PCR, LaFountian et al. TWGDAM Validation of the AmpFlSTR Profiler Plus and the AmpFlSTR Cofiler STR Multiplex Systems Using Capillary Electrophoresis 2001 J Forensic Sci 46(5):1191-1198. The two STR kits were validated in a forensic lab with a variety of sample types from purified DNA to blood on FTA to complex sample types (blood on blue denim). Many parameters were tested both PCR and capillary electrophoresis. DNA from blood on FTA performed as well as genomic DNA purified by proteinase K and organic extraction. forensic, validation experiments, human ID Lam PF and Shaari A Rapid PCR Screening of Transgenic Pineapple using FTA Cards 2003 Poster: 14th Malaysian Society of Plant Physiology Conference, Awana Genting, Malaysia Leaves of transgenic and non-transgenic pineapple were crushed onto FTA cards and examined for the presence of the transgene by PCR transgenic plants, plant, agriculture FTA Reference DatabaseJun05 15

Lampel et al. Improved Template Preparation for PCR- Based Assays for Detection of Food-Borne Bacterial Pathogens 2000 Applied and Environmental Microbiology 66: 4539-4542 FTA based technique for the detection of food microbes in wash solutions for produce & beef. Shigella and Salmonella are gram-negative easy to lyse and fewer are detectable on FTA; Listeria are grampositive harder to lyse need more to detect on FTA. Shigella flexneri, Salmonella enterica, Listeria monocytogenes, gram-negative, grampositive, produce, wash procedure, ground beef, boiling, food safety, screening, Lampel KA & Orlandi PA Rapid detection of pathogenic microbes by PCR using a nonextraction template protocol 2002 Poster IUMS International Conference, Paris The inherent properties of FTA filters allows PCR template preparation for microbial organisms Lampel,KA, Dyer Detection of Bacillus Spores Using D, Kornegay L PCR and FTA Filters. and Orlandi PA Lampel et al. Real-Time PCR Detection of Shigella From Foods Using FTA Filters. Lappas et al. The utility of the FTA GeneCard system for analysis of buccal swab transfers Ledray & Netzel Forensic Nursing: DNA Evidence Collection 2004 J Food Protection 67(5)1036- Difficult to lyse spores from Bacillus subspecies 1038. 2004 Poster: 104th General Meeting American Society of Microbiology, New Orleans 1999 Poster 10th International Symposium on Human ID 1997 Journal of Emergency Nursing 23:156-158 cereus, thruingiensis, subtilis and niger were isolated and applied to FTA cards. A fragment of the 16S rrna gene was amplified by PCR. Sensitivity of approx 50 spores were detected and this was increased to 5 spores by nested PCR. Shigella flexneri were added ("spiked") to apple cider, strawberries or cilantro. PBS (10ml) was used to wash berries and cilantro. An 10ul aliquot of food wash or spiked cider was applied to FTA. After sample application the filters were washed with ethanol then dried at 56 o C. The filters were then washed twice with FTA reagent then twice with TE-1 and dried at 56 o C. Disks of 6mm were cut out and added to 50ul TE and heated at 95 o C for 10 min to elute DNA for real-time PCR. Nested real-time PCR done on a LightCycler with SYBR Green incorporation and by hybridization probe displacement. Both real-time PCR methods can be successfully used with FTA. Using FTA a sensitivity level of less than 10 organisms cam be achieved from pure culture or seeded foods. FTA solves the problem of DNA extraction & shipping issues. Describes use of FTA in blood collection and storage from victim and/or perpetrator in rape cases bacteria, environmental, food safety, bacteria, pathogens, food borne illness, food safety, detection, nested PCR 150bp, >12Kb, stable DNA, intact DNA blood, victim, databasing FTA Reference DatabaseJun05 16

Li et al. Persistence of Human Immunodeficiency Virus Type 1 Subtype B DNA in Dried-Blood Samples on FTA Filter Paper 2004 J. Clin. Microbiol. 42(8)3847- HIV-1 DNA is stable on FTA cards for over 4 years 3849. from time of collection. Previous data only showed stability on untreated paper as long as 15 months. HIV, DNA stability Lim SES and Tan WF Evaluation and Modification of the IsoCode Card DNA Isolation Method 2002 Poster: 16th Meeting of the International Assocication of Forensic Science, Montpellier France, September Blood was applied to FTA and to IsoCode. IsoCode yields amplifiable DNA and is comparable to the results achieved with their normal methods using FTA IsoCode, blood Lin et al. Detection of Plant Genes Using a Rapid, Nonorganic DNA Purification Method 2000 BioTechniques 28: 346-350 FTA used to process PCR ready template from a wide variety of plant samples Arabidopsis, marijuana, Cannabis, cassava, coca, corn, orchid, papaya, petunia, opium poppy, soybean, sugarbeet, sugarcane, tobacco, tomato, transgenic, uida gene, CTAB, plant DNAzol, 18S rrna gene, rbcl gene, soybean cyst nematode race 3 resistance gene, monocot, dicot, marker assisted breeding, selection, Littlejohn et al. Determination of b2-adrenergic Receptor (ADRB2) Haplotypes by a Multiplexed Polymerase Chain Reaction Assay. MacLean L, Chisi Severity of Human African JE, Odiit M, Trypanosomiasis in East Africa is Gibson WC, Associated with Geographic Location, Ferris V, Picozzi Parasite Genotyp, and Host K and Sternberg Inflammatory Cytoking Response JM Profile 2002 Human Mutation Mutation in Brief #562, Online. 2004 Infection and Immunity 72(12):7040-7044 Peripheral blood was either applied to FTA cards or DNA ARMS assay, SNP, extracted by the following methods, guanidine multiplex, clinical study isothiocyanate, sodium hydroxide boiling lysis or salting out. FTA punches or extracted DNA was then analyzed in a multiplex PCR assay to detect SNPs in the ADRB2 gene. Results showed that FTA was not as efficient in this study as DNA extracted by the other methods. During surveillance for typanosomaisis, 200ul of a buffy coat suspension (primarily white blood cells) were applied to FTA cards which where then dried at room temperature to detect trypanosome DNA by PCR amplification of 2mm punches protozoan, protozoa, population study, disease detection FTA Reference DatabaseJun05 17

Martinez- Gonzalez et al. Biological Stains Collected from Crime Scenes using FTA Paper. 2005 Poster: 57th Annual Meeting of the American Association of Forensic Scientists, New Orleans LA. FTA was used to blot blood and semen stains from nonabsorbent (glass, tile, wood) and from absorbent (carpet) surfaces. First wet the stain with distilled water then blot with FTA cards. This method is quick, easy and preserves the original stain since only some of the cells are transferred. This method is very applicable to analysis of biological stains at crime scenes. crime scenes, sample recovery Martins et al. Haplotype Study of Microsatellites Flanking the t(15;17) Breakpoint in Acute Promyelocytic Leukemia Patients from North Portugal 2002 Leukemia 16:1353-1357. FTA used to collect blood from unrelated, healthy individuals and blood from PML patients and their families. DNA was extracted from FTA using chelex. human, blood, acute promyelocytic leukemia, PML, microsatellites, PCR, eluted DNA, chelex Matsumoto et al. Tiburonia granrojo n. sp., A Mesopelagic Scyphomedusa from the Pacific Ocean Representing the Type of a New Subfamily (class 2003 Marine Biology 10.1007/s002227-003-1047-2 (online journal) FTA used in the identification of a new jellyfish species. Tissue was pressed on to FTA which then was PCR amplified. jellyfish, tissue, 28S rrna gene, PCR, sequencing, new species, genotyping Scyphozoa: order Semaeostomeae: family Ulmaridae: subfamily Meadus and MacInnis Tiburoniinae subfam.nov.) Simplifying Genetic Testing from Porcine Hair and Blood Samples 2000 Presentation: Banff Pork Seminar FTA used to collect blood from ear vein prick of porcine livestock. non-invasive, genetic testing, pigs, Mills et al. Effect of Immunosuppression on Outcome Measures in a Model of Rat Limbal Transplantation. 2002 Invest Opthalmol Vis Sci. 43(3)647-655 FTA was used to collect cells from coreal transplants in rats to identify the presence of donor cells by PCR. eye tissue, transplantation, tissue grafts, limbal tissue Moran et al. High Throughput Purification of Samples Archived on FTA Cards 2003 Poster:14th International Symposium on Human ID, Shows the use of BioMek 2000 for washing punches from blood, buccal and plant samples Automation of FTA Phoenix AZ Moran et al. The use of FTA for the Identification of Bacteria from Culture Collections and Clinical Isolates. 2004 Poster: 104th General Meeting of the American Society of Microbiology, New Orleans LA. Demonstrates use of FTA for characterizing bacterial genomic DNA samples. PCR amplification of species specific genes and RAPD analysis. Some gram positive species of Streptococcus and Peptostreptococcus were used in this study. Bacterial ID, environmental, clinical, microbiology, gram positive Moran et al. Comparison of DNA Stability on Treated and Untreated Papers. 2004 Poster: 56th Annual Meeting of the American Association of Forensic Scientists, Dallas TX Shows studies of degradation of DNA on untreated filter paper compared to preserved DNA on FTA. Accelerated aging by UV radiation of the DNA showed that FTA is superior to untreated filter papers for preserving the integrity of DNA. UV radiation, aging, DNA stability FTA Reference DatabaseJun05 18

Moran B and Layton R Analysis of Cattle Samples on FTA Cards using Stockmarks Genotyping Kit 2005 Poster: XIII Annual Plant and Animal Genome Conference, San Diego CA Blood and buccal samples were collected from Holstein cattle, animal ID, cows and applied to FTA cards. Samples were analyzed genotyping, SNP analysis by the ABI Stockmarks genotyping kit which employs microsatellite markers and also by SNP analysis. The data were very similar for both analytical methods although the SNP analysis could analyze more difficult samples. FTA proves to be useful for collecting cattle samples for both PCR and SNP analysis. Morton et al. A Simple Heat Elution of Human Genomic DNA from FTA Treated Paper Using Water 2000 Poster: 11th International Symposium on Human ID FTA benefits of destroying potentially harmful pathogens and long term room temperature storage. Heated water eluted enough DNA to quantify and be used in downstream forensic analysis. eluted DNA, GenSpin, quantitate DNA, DNA in solution, Moscoso et al. PCR Detection of Mycoplasma Stored and Transported on FTA Gene Guard Filter Paper 2003 Poster: World Vet. Poultry Assn & 140th AVMA Annual Convention, Denver CO Used FTA to detect two subspecies of mycoplasma. Fluid viral inactivation, containing live infectious bronchitis virus and Newcastle pathogen inactivation, Disease virus were inactivated after 1 hr on FTA. Also chicken, diagnostic, both species of mycoplasma were inactivated after being animal biotechnology spotted onto FTA. Both purified cultures and field collected samples from chickens were spotted onto FTA and analyzed by PCR for the 16S rrna gene. Moscoso et al. Molecular Identification of Avian RNA Viruses Stored on FTA Filter Paper. 2004 Poster: 53rd Western Poultry Disease Conference, Sacramento CA. RNA viruses Infectious Bronchitis Virus, Infectious Bursal Disease virus and Avian Leukosis Virus (subgroup J) from chicken samples were applied to FTA. FTA inactivated the viruses. RNA was extracted from the cards and was analyzed by RT-PCR. RT-PCR was specific for the target virus. RT-PCR from bursa on FTA can detect as little as 5 ng of RNA. RNA is stable on FTA cards. chicken, bursa, tracheal swab, allantoid fluid, tumors, liver, RFLP, strain differentiation, pathogen inactivation Moscoso et al. Indentification of Infectious Bronchitis Virus by RT-PCR from FTA Filter Paper 2004 Poster: International Poultry Scientific Forum, Atlanta GA FTA was used to collect and store 5ul aliquotes of allantoic fluid containing live virus. Cards were stored at room temp, 4 o C and 41 o C before analysis. Two mm disks were washed in TE-1, vortexed and incubated at room temp 15 min. RNA was extracted using High Pure Viral RNA Kit (Roche). RNA served as a template for RT-PCR and RFLP. Live virus was inactivated after application to FTA. FTA was sensitive for RNA detection. RNA on FTA stored at 41 o C for 15 days still showed good RT-PCR demonstrating the protectective effect of FTA. viral inactivation, pathogen inactivation, chicken, diagnostic, animal biotechnology, RNA stability Moscoso H, Thayer SG, and Hofacre CL Application of the FTA Technology for the Collection, Shipment and Processing of Avian DNA and RNA Viruses 2004 Poster: 141st AVMA Annual Convention Philadelphia, July Adenovirus and ILTV are DNA viruses, New Castle Disease virus is ssrna virus. These were detected using samples of liver, tracheal swab or tissue culture samples applied to FTA. RNA was eluted from the disks in Tris EDTA buffer, extracted then amplified by RT-PCR for NDV. As little as 5 ng of RNA virus was detected on FTA. disease diagnostics, DNA virus, RNA virus, inactivation FTA Reference DatabaseJun05 19

Moscoso et al. Inactivation, Storage and PCR Detection of Mycoplasma on FTA Filter Paper 2004 Avian Diseases 48:841-850. FTA was used to detect two species of Mycoplasma in pure cultures and in field specimens. 193 field samples were spotted to FTA, analyzed by PCR and the results compared to classical serology methods of detection. Nearly 100% agreement occurred between FTA and classical methods. FTA completely inactivated the mycoplasma and serves as a safe and cost effective method to transport samples to the diagnostic lab. disease diagnostics, avian disease, tissue samples, tracheal swabs, Moscoso et al. Moscoso et al. Moscoso et al. Molecular Detection and Serotyping of Infectious Bronchitis Virus from FTA Filter Paper Molecular Characterization of Fowl Adenoviruses from FTA - Liver Impressions Detection of Infectious laryngotracheitis Virus by PCR on Specimens Stored on FTA Filter Paper 2005 Avian Diseases 4 9:24-29 Infectious Bronchitis Virus (IBV) is an RNA virus. RT-PCR assays were devloped to detect IBV and variants. Virus propagated in allantoic fluid or tracheal swabs from chickens were applied to FTA cards. Tests showed that virus applied to FTA cards was inactivated the virus after application to the cards making international shipment of test samples to the diagnostic lab easy. RNA was extracted from the FTA disks using the High Pure Viral RNA Kit (Roche) or using Trizol. RNA was stable on FTA with only a slight decrease detected after 15 days at 41 o C. 2005 Poster: 54th Western Poultry Inclusion body hepatitis (IBH) is caused by adenoviruse Disease Conference. Vancouver Canada April 25-27 group I and can now be diagnosed by molecular techniques. Sections of infected liver was applied to FTA by squashing the tissue onto the card. Adenovirus was shown to be inactivated. Several serotypes of virus were detected by PCR after 8 months of storage on FTA. Sequencing of amplicons showed 100% homology to the challenging virus so no change in sequence occured after storage on FTA. This is the first time adenovirus was detected from liver impressions on FTA cards. 2005 Poster: 54th Western Poultry Infectious Laaryngotracheitis viral (ILTV) disease had Disease Conference. Vancouver Canada April 25-27 been detected by histopahtology or direct antibody detetion taking over a day to diagnose. Molecular detection of the disease on FTA has greatly speeded up the diagnosis process. Sample types on FTA included eyelid impressions, trachea impressions, tracheal scrapings and tracheal swabs. ILVT was inactivated on FTA. ILTV was detected from FTA by PCR. Eyelid impressions served to be the best souce of sample for FTA cards and 100% successful diagnostics. virus, RT-PCR, RNA virus, viral inactivation, veterinary diagnostics, RNA stability, DNA virus, liver samples, tissue, viral inactivation, sequencing DNA virus, conjunctivitis, viral inactivation FTA Reference DatabaseJun05 20

Nakayama et al. Clinical Significance of Circulating DNA Microsatellite Markers in Plasma of Melanoma Patients 2000 Ann NY Acad Sci 906; 87-98 FTA used to collect blood from patients with melanoma Loss of heterozygosity, human, cancer study, clinical, diagnostics Natarajan et al. Paper-Based Archiving of Mammalian and Plant Samples for RNA Analysis 2000 BioTechniques 29: 1328-1333 FTA used for storing & analyzing RT-PCR mrna from plant & tissue culture cells- several months ambient storage (longer -70 C) 2mm punch for RT-PCR, HeLa cells, BHK-21 cells, cell culture, blood, buccal, plant, leaf, potato, poly(a)+ mrna, total RNA, Trizol, Northern blot, Ndunguru et al. Application of FTA Technology for Sampling, Recovery and Molecular Characterization of Viral Pathogens and Virus-derived Transgenes from Plant Tissue. 2005 Virology J 2:45 Both DNA and RNA viruses were detected in several plant species after application to FTA. The FTA method is comparable to tradtional methods of recovering RNA and DNA viruses and FTA simplifies the sampling and analysis of diseases plants in both the lab and the field. Amplified sections of DNA were cloned into sequencing vectors and the sequences compared at the 99.8% level to DNA purified by traditional methods geminiviruses, maize, cassava, tomato, tobacco, Tobacco mosaic virus, Potato Virus Y and Tobacco etch virus, ssdna virus, East African cassava mosaic Cameroon virus, African cassava mosaic viurs, Tomato yellow leaf curl virus Neimark et al. Mycoplasma ovis comb. Nov. (formerly Eperythrozoon ovis), an epierythrocytic agent of haemolytic 2004 Int J Systm Evol Microbiol. 54:365-371. Blood samples from sheep were deposited on FTA. The 16S rrna gene of a red cell parasite was amplified by PCR. The results showed that the parasite though to bacteria, pathogens, animal biotechnology, mycoplasma anaemia in sheep and goats. be a member of Rickettsia is acutally a member of the mycoplasma family of pathogens. Neimark et al. Phylogenetic analysis and description of Eperythrozoon coccoides, proposal to transfer to the genus Mycoplasma as Mycoplasma coccoides comb. nov. and Request for 2005 Intl J Systematic Evol Microbiol. 55:1385-1391 Blood from mice infected with E. coccoides was applied to FTA cards. Three mm punches were washed with FTA purification reagent and TE (not TE-1) and used as template for 16S rrna gene PCR. Amplicons were sequenced and sequences compared to other species of Mycoplasma. phylogenetic tree construction, blood parasites, Nelson et al. Opinion Detection of a Primer-Binding Site Polymorphism for the STR Locus D16S539 Using the PowerPlex 1.1 System and Validation of a 2002 Journal of Forensic Science 47 (2): 345-349 DNA database sample discrepancies were caused by the amplification conditions (AmpliTaq vs. AmpliTaq Gold) when using Promega's PowerPlex 1.1 typing kit and not by the different DNA extraction methods (FTA vs. organic extraction). allele dropout, hotstart PCR, DNA typing, point mutation, polymorphism, imbalanced alleles, Degenerate Primer to Correct for Polymorphism FTA Reference DatabaseJun05 21

Nields et al. Evaluation of the FTA DNA Collection and Storage System on Autopsy Blood from Medical Examiner Cases 1998 Poster American Academy of Forensic Sciences FTA for blood processing of human autopsy cases. Samples taken up to 12 hr after death. medical examiner, autopsy, Norton and Shriver O'Shea et al. Orlandi & Lampel Orlandi et al. Orlandi and Lampel Whole Genome Amplification on FTA Cards. Amplified Fragment Length Polymorphism Reveals Genomic Variability among Mycobacterium avium subsp. paratuberculosis Isolates. Extraction Free, Filter-based Template Preparation for Rapid and Sensitive PCR Detection of Pathogenic Parasitic Protozoa Targeting Single Nucleotide Polymorphisms in the 18S rrna Gene to Differentiate Cyclospora species and Eimeria species by Multiplex PCR. FTA Filter Applications: PCR Format for Alleviating Technical Barriers in the Detection of Food-borne Pathogens in Complex Matrices. 2003 Poster: IX Plant and Animal Genome Conference, San Diego, USA DNA from blood spots collected on FTA was used as a template in Whole Genome Amplification (WGA). Punches were washed using a Hydra liquid handler and amplified using a Templiphi kit (Amersham). After WGA, 15 different loci were amplified for analysis with average success rates of 90-96%. 2004 J. Clin. Microbiol. 42(8) 3600-M. avium subsp. paratuberculosis causes Johne's 3606. disease a debilitating disease in cattle. Twenty isolates of this pathogen were spotted to FTA cards. FTA punches were washed according to the standard protocol then 7.1µl of ddh 2 O were added and the punch heated to 94 o C for 5 min, 12.9µl of master mix were 2000 Journal Clinical Microbiology added for PCR amplification. FTA used to isolate protozoan oocyte DNA from soft fruit 38: 2271-2277 and stool - sensitive detection and time-saving procedure. 2003 Appl. Envron. Microbiol. 69:4806-4813. Parasite oocytes were applied to FTA to prepare DNA template for both nested PCR or PCR followed by RFLP to differentiate species of Cyclospora and Eimeria. Raw fecal material was spotted to FTA and subjected to SNP Multiplex PCR. This method quickly identifies the species designation of clinical isolates of parasites. rolling circle amplification, Phi DNA polymerase, multiple strand displacement, MDA, gram positive bacteria, pathogen, cattle, clinical diagnostics parasites, Cyclospora, Cryptosporidium, microsporidia, Encephalitozoon intestinalis, spores, multiplex PCR, food, safety, screening, fecal specimens, feces, Parasite, fecal, feces, RFLP, SNP, multiplex, species specific, environmental 2003 BioProcessing J 2(6)45-54. Technology review of the current methods of detecting food-parasitesborne pathogens vs using FTA as a faster and simpler bacteria, pathogens, food microbes, method for preparing DNA for analysis. Current methods safety, environmental of growth enrichment or selection are not necessary using the FTA method thus shortening the prep time dramatically. FTA Reference DatabaseJun05 22

Patel AA PCR Detection of Streptococcus Mutans and Streptococcus Sobrinus in Dental Plaque Samples from Low, Moderate and High Caries Risk Children 2002 MA Thesis Virginia Commonwealth University Plaque samples were taken from volunteers with a sterile toothpick and placed into 25ul bufffer and frozen. After thawing 10ul were spotted to FTA cards. A 2mm punch was taken and washed as normal. A 50ul PCR master mix was added to the punches and amplified for the specific bacteria under examination. bacterial id, clinical microbiology, disease diagnostics, human, dental disease Prager et al. Picozzi et al. Chlamydia pneumoniae in Carotid Artery Atherosclerosis A Comparison of its Presence in Atherosclerotic Plaque, Healthy Vessels, and Circulating Leukocytes from the Same Individuals. The diagnosis of trypanosome infections: applications of novel technology for reducing disease risk. 2002 Stroke 33:2756-2761. Whole blood collected from patients with atherosclerosis and DNA prepared by Miller salting out method and also spotted to FTA. No difference was seen between DNA isolated by either method when analyzed with PCR for the presence of the infectious bacterial pathogen Chlamydia pneumoniae. The 16S rrna gene fragment was amplified for identification of the pathogen. 2002 African Journal of Biotechnology 1 :39-45 PCR on buffy coat preparations using Whatman FTA matrices were the gold standard in the field of sleeping sickness. human, bacteria, pathogen, diagnostic, microbes, parasites, cattle, trypanosome, Trypanosoma brucei rhodesiense, Trypanosoma brucei brucei, T. vivax, T. congolense, livestock, screening, animal health, field testing, Picozzi et al. Diagnosis of Trypanosoma brucei infections in cattle: applications of novel technology for estimating disease risk. 2003 ICPTV Newsletter 8 pg 24-25 September 2003 Comparison of DNAzol, whole blood on FTA and buffy coat on FTA to prepare DNA from cattle for screening of the trypanosome parasite that causes sleeping sickness in humans. The parasites were detected by PCR analysis of the DNA. Advantages of the FTA are easy field collection and the ability to re-analyze samples due to the long storage capacity of FTA cards parasite, buffy coat, whole blood, cattle, disease, diagnosis, PCR, field collection, storage Pierce et al. High Throughput DNA Purification of Samples Archived on FTA Cards. 2002 Poster: 12th Genome Sequencing and Analysis Conference, Boston MA Demonstrates the use of Biomek 2000 in washing FTA punches prior to PCR analysis. automation, liquid handling, blood, plant, buccal, STR, Raina & Dogra Application of DNA Fingerprinting in Medicolegal Practice 2002 J. Indian Med. Assoc. 100(12) 688-694 Use of FTA for blood sample collection, archive and DNA preparation for PCR based DNA fingerprinting (STR analysis). Mentions features and benefits of FTA, i.e., easy, can archive samples at room temp, no quantitation is necessary from the punch and punch can be reused in another PCR. microsatellites, biological samples FTA Reference DatabaseJun05 23

Rajendram et al. Microbial Applications of FTA Technology 2002 Poster IUMS International Conference, Paris Bacterial strains and clinical isolates stored on FTA. Ribotyping of bacteria with 16s rrna gene done successfully on gram positive and gram negative bacteria. Not all spores were lysed and inactivated. Corynebacterium, Nocardia, Mycobacterium, Bacillus, Neisseria, Staphylococcus, Streptococcus, Enterococcus, Bordetella, Legionella, Pseudomonas, Pasterurella, Aeromonas, Haemophilus, Shigella, Citrobacter, Proteus, Moraxella, vibrio, Actinobacillus, Peptostreptococcus Rajendram DNA Storage and Recovery for DNA Fingerprinting and Gene Detection using FTA Paper Technology 2003 Presentation PHLS/Whatman symposium, London 400 strains of bacteria tested on FTA; gram-negative, gram-positive and spore formers. 16S rrna gene amplified by PCR as well as single copy genes showing sensitivity of FTA DNA prep. Archive bacterial DNA on FTA for up to 1 yr. AFLP and RAPD analysis done. FTA inactivates bacteria at low to moderate titer. At high titer some gram-positive are viable. Additional lysis with guanidine thiocyanate inactivates all bacteria. corynebacterium, Nocardia, Mycobacterium, Bacillus, Neisseria, Staphylococcus, Streptococcus, Enterococcus, Bordetella, Legionella, Pseudomonas, Pasterurella, Aeromonas, Haemophilus, Shigella, Citrobacter, Proteus, Moraxella, vibrio, Actinobacillus, Peptostreptococcus, Clostridium, ribotyping Rejt et al. Genetic Variability of Urban Kestrals in Warsaw - Preliminary Data 2004 Zool. Poloniae 49: 199-209 Kestrals are birds of prey like hawks. Blood samples from the birds were collected on FTA cards. Punches, 2mm, were included in 25µl reactions to amplify microsatellite markers. Birds, population study, ecology, population diversity FTA Reference DatabaseJun05 24

Rensen et al. Devlopment and Evaluation of a Real- Time Fluorescent Polymerase Chain Reaction Assay for the Detection of Bovine Contaminates in Cattle Feed. 2005 Foodborne Pathogens and Disease 2(2)152-159 FTA was used to detect Bovine Meat and Bone Meal (BMBM) contaminants in two kinds of cattle feed. 10gram samples of BMBM spiked feed were extracted with a lysis buffer and after extraction the lysis buffer was spotted to FTA cards. Disks, 2mm, were punched and treated with DNA-free RNase to increase the sensitivity of the PCR. The DNA was extracted from the punch using Instagene (Chelex) matrix. Extracted DNA was used in real time PCR amplifications. The authors say this method employing FTA can be easily automated for routine screening. cattle feed, bovne, contaminants, ruminant protein, runinant byproducts Renshaw et al. Roberts et al. Amplification of DNA from blood spots: the importance of collection paper & primer sequence Rapid and Comprehensive Determination of Cytochrome P450 CYP2D6 Poor Metabolizer Genotypes by Multiplex Polymerase Chain Reaction. 2001 Focus 23: 8-9 The yield of PCR product using DNA on FTA as a template was twice as high as when using DNA on Guthrie cards as a template for Factor V amplification. 2000 Human Mutation 16:77-85. Peripheral blood was either applied to FTA cards or DNA extracted by the following methods, guanidine isothiocyanate, sodium hydroxide boiling lysis, proteinase K/phenol-chloroform or salting out. FTA punches or extracted DNA was then analyzed in a multiplex PCR assay to detect different forms (alleles) of the CYP2D6 gene. The ARMS assay used here is a multiplex PCR assay to detect several alleles at a time and is conducted from a >4Kb DNA fragment amplified from genomic DNA. FTA was a suitable template to generate the >4Kb starting material for the ARMS assay. The authors found that a hotstart PCR was essential when using FTA punches. Factor V Leiden, methylenetetrahydrofilate reductase, MTHFR, polymorphism, thromboembolic, disease, Guthrie cards, blood, human, infant, ARMS assay, SNP, multiplex, clinical study, long PCR, alleles, hotstart PCR. Rogers & Burgoyne Reverse Transcription of an RNA Genome from Databasing Paper (FTA) 2000 Biotechnology Applied Biochemistry 31:219-224 Archiving of RNA genomes at room temperature using FTA. Detecting RNA in transfusion blood samples. Different methods tried to keep RNA on filter. Coxsackievirus, CVB-4, RT-PCR, virus, diagnostics, immobilized RNA, viral, transport, safety, enterovirus, FTA Reference DatabaseJun05 25

Rogers & Burgoyne Bacterial Typing: Storing and Processing of Stabilized Reference Bacteria for PCR Without Preparing DNA-An Example of an Automated Procedure 1997 Analytical Biochemistry 247: 223-227 Illustrates FTA for collecting, storing, rapidly analyze microorganisms. 16S, 23S, ribosomal RNA genes, archiving, ribotyping, Staphylococcus, E. coli, automation, genetic ID, infections, pathogenic, clinical isolates, Rogers et al. Development of a Buccal Swab Kit for the Collection of DNA Database Samples 1997 Poster: 8th Annual Int. Symposium of Human Identification Scottsdale, AZ, USA Use of a buccal collection kit for DNA analysis. offender specimens, buccal, foam tipped applicator, training Rogers et al. Implementation of a Buccal Swab Kit for the Collection of DNA Database Samples 1998 Poster: 9th Annual Int. Symp. on Human Identification Orlando, FL, USA Buccal sample processing on FTA for STR analysis- Potential PCR inhibitors in saliva don't reduce efficiency of PCR on FTA. DNA database, buccal, DNA typing, contaminants, training, beverages, candy, tobacco, food, lipstick, mouthwash, Salvador & De Ungria Isolation of DNA from Saliva of Betel Quid Chewers Using Treated Cards. 2003 J Forensic Sci 48(4)749-797 FTA used to collect buccal cells for DNA analysis from people who chew betel quid. Betel quid contains DNA damaging agents and PCR inhibitors. FTA very successfully isolated the DNA away from the agents and inhibitors to allow PCR amplification. plant, human, polyphenolics, PCR inhibitors, STR, field collection, genetic studies Salvador JM and MCA De Ungria DNA Stability of Forensic STR Markers in FTA Extracted Buccal DNA of Betel-quid Chewers. 2004 Oral Oncology 40:231-232. Buccal cell DNA from betel quid chewers may be unstable as determined by dropout of STR loci when cell samples are collected by traditional methods and DNA isolated by organic extraction. Buccal cells collected and applied to FTA did not show this instability suggesting that the organic extracted DNA contained an agent that degraded the DNA. buccal cells, genetic ID, human ID, plant extract Sauerbrei et al. Genetic Profile of an Oka Varicella Vaccine Virus Variant Isolated from an Infant with Zoster. 2004 J. Clin. Microbiol. 42(12)35604-5608. Varicella virus causes chickenpox. People vaccinated multiple PCRs, pure DNA against chicken pox with an inactivated viral solution can stored on FTA, viral DNA sometimes come down with shingles (zoster). DNA from detection, DNA viruses blood samples was purified with Qiagen Easy DNA kit or QIAamp blood kit. Purified DNA was stored on FTA at room temp for up to 1 month before PCR. 4mm squares of FTA were washed as normal and used at least three times in PCRs for multiple SNP detections. Approx 100 ng of DNA were on the filter pieces. FTA Reference DatabaseJun05 26

Schifferli et al. DNA Obtention to Study BLAD in Argentine and Spanish Cattle. 2000 Arch. Zootec. 49:505-508. (in spanish!) Animal biotechnology, blood, diagnostic, breeding stock. Schulman et al. Microsatellite Analysis of Cryopreserved Stallion Semen Stored on FTA Paper. 2002 J South African Vet Assoc 73(4):222-223. Use of FTA to sample sperm and blood from stallions for horse, frozen, blood, genetic ID matching. Processed sperm (concentrated by sperm, microsatellites, centrifugation and most seminal fluid removed) diluted 1:10 genetic screening, or 1:100 put on FTA. DDT and Proteinase K added to FTA artificial insemination purification reagent to lyse sperm before multiplex PCR. Seabury, CM and JN Derr Identification of a novel ovine PrP polymorphism and scrapie-resistant genotypes for St. Croix White and a related composite breed. 2003 Cytogenet Genome Res 102:85-88. Whole blood collected from sheep onto FTA for PCR analysis and the amplified DNA was sequenced to look for DNA base changes. A 1.2mm punch was used in a 25ml PCR. A new DNA polymorphism may help to determine if some sheep are resistant to developing scrapie. sheep, blood, animal biotechnology, polymorphism, disease resistance Seah & Burgoyne DNA Archiving on FTA Paper Photosensitizer Initiated Attacks as Models of Aging 2000 Poster 4th HUGO Pacific Meeting and 5th Asia-Pacific Conference on Human Genetics Describes microgel electrophoresis of plasmid DNA from FTA. Studies of FTA stabilizing DNA in accelerated aging treatments. DNA stability, aging, strand breaks, riboflavin, hematoporphyrin, free radicals, Seah & Burgoyne DNA Databasing on FTA Paper: Biological Assault and Techniques for Measuring Photogenic Damage 2000 Progress in Forensic Genetics 8:74-77 Describes the microgel electrophoresis technique to measure integrity of DNA on FTA after attempted damage from accelerated ageing (free radicals, UV and mould attack) DNA stability, aging, strand breaks, riboflavin, hematoporphyrin, free radicals, humid conditions, Seah & Burgoyne DNA Databasing on FTA Paper: Biological Assault and Techniques for Measuring Photogenic Damage 2000 Poster International Society for Forensic Haemogenetics 1193 Illustrates the ability of FTA to prevent damage of stored nucleic acid by UV DNA stability, aging, strand breaks, riboflavin, hematoporphyrin, free radicals, humid conditions, Seah & Burgoyne The Study of DNA Integrity on Databasing Material: an In-situ Electrophoretic Method 1998 Poster 14th International Symposium on Forensic Sciences. The Francois Vidocq Conf Oct 1998. Used an in-situ electrophoretic technique to determine DNA damage over time, and under different storage conditions for DNA stored on plain paper and FTA Cards. The DNA on FTA showed less damage and no fungal growth when compared to plain paper. DNA stability, high humidity, ambient storage, blood, buccal, FTA Reference DatabaseJun05 27

Seah & Burgoyne Photosensitizer initiated Attacks on DNA Under Dry Conditions and their Inhibition: a DNA Archiving Issue. 2001 J. Photochem and Photobiol B: Biology 61: 10-20. Free radical reactive ions which normally damage DNA were artificially generated to "damage" plasmid DNA on FTA. This paper studies how FTA protects DNA from the damage and tries to quantitate the damage using microgel electrophoresis. DNA damage, dry DNA Seah et al. da Silva et al. da Silva et al. Sipes et al. STR Data for the AmpFL STR Identifiler loci in three ethnic groups (Malay, Chinese, Indian) of the Malasian population. Visceral leishmaniasis Caused by Leishmania (Viannia) braziliensis in a Patient Infected with Human Immunodeficiency Virus. Diagnosis of Human Visceral Leishmaniasis by PCR using Blood Samples Spotted on Filter Paper FTA Beyond the Everyday: Novel FTA Applications to Improve Efficiency. 2003 Forensic Science International 138:134-137. Blood was collected from 638 people onto FTA. Punches (1.2mm) were taken and washed. The 15 STR loci in the Identifiler kit were amplified directly from the FTA punch. 2002 Rev. Inst. Med. Trop. S. Paulo Patient blood and bone marrow collected onto FTA. 44(3) 145-149. 2004 Genetics and Molecular Res. 3(2): 251-257 2000 Poster: 11th International Symposium on Human ID After washing punch containg purified DNA was amplified by PCR to detect Leishmania. Reference strains were applied to FTA and purified DNA amplified as positive controls. Blood and bone marrow aspirates as a volume of 30ul was applied to FTA cards and washed as usual prior to PCR amplification. Using molecular techniques visceral leishmaniasis can be diagnosed instead of using microscopic examination or immunoassays for the blood, human id, allele frequencies, population study parasite, co-infections, blood, bone marrow, diagnostic, diagnosis, clinical samples, environmental parasites, bone marrow, evidence of parasites. DNA stored on non-fta coated paper can be processed throughput, DNA extract, for amplification using the FTA procedure but high MW static charge, DNA is degraded and the yield is low. Freezing punches in FTA Purification Reagent gave poor results. It is not necessary to let the punch dry completely after the last wash but all wash must be removed. Sippel & Sajantila Forensic Casework Experience: FTA Bloodstain Card as a Matrix for Blood Samples from Fresh and Decomposed Bodies. 2000 Poster Promega 11th Annual Int Symp Of Human ID. FTA is useful in collecting samples from autopsy cases decomposition, STR, QiAquick, human ID Siripattanapipon Genotype Study of Pneumosystis g et al. jirovecii in Human Immunodeficiency Virus-positive Patients in Thailand 2005 J Clin Microbiol 43(5):2104-2110. Pneumocystis jirovecci is a fungus causing pneumonia in immunosuppressed individuals. Approx 14ml of bronchioalveolar lavage or sputum was applied to a 6 mm disk of FTA. One fourth of the disk was washed with FTA purification reagent followed by TE-1 then dried at 80 o C. The washed FTA was used directly in a 50ml PCR amplification. Fungus, clinical samples, disease diagnostics. FTA Reference DatabaseJun05 28

Sitaraman et al. Amplification of Large DNA from Blood Stored at Room Temperature 1999 Focus 21(1):10 PCR fragments of 4.1, 5.2, 7.5, 8.0 and 8.4 kb amplified from FTA punches containing blood human, blood, large PCR fragments, Taq, Platinum Taq, Platinum Taq High Fidelity Smith & Burgoyne Species identity: conserved inverted LINE repeat clusters (ILRC) in the vertebrate genome as indicators of population boundaries 2001 Gene 271: 273-283 Species identity study used FTA paper to collect, ship, store and analyze DNA from several species of birds, mammals and amphibians. DNA from birds and lizard was heat eluted and soluble DNA used in PCR. blood, birds, chicken, cat, horse, cow, rat, lizard, tissue, PCR, cloning, DNA sequencing, Smith, LM and Burgoyne, LA Collecting, archiving and processing DNA from wildlife samples using FTA databasing paper. 2004 BMC Ecology 2004, 4:4 (Article available at http://www.biomedcentral.com/ 1472-6785/4/4). A variety of wildlife sample types were collected and processed on FTA for PCR analysis. Blood samples, blood clots, tissue, saliva and buccal cells samples were applied to FTA. Several variations of processing FTA disks are described. Techniques for nucleated avian red blood cells are described. Methods to elute DNA from FTA are described. horse, chicken, cow, crested pigeon (Geophaps lophotes) and sleepy lizard (Tiliqua rugiosa), pelicans, mallee fowl (Leipoa ocellata), frog (Rana sp.), yabbie (an Australian fresh water crustacean, Cherax tenuimanus ), abalone (Haliotis laevigata ) and blue swimmer crab (Portunus pelagicus), King George Whiting (Sillaginodes punctata ) and tuna (Thunnus maccoyii) Smith et al. FTA Technology New Advances for the Collection, Shipment, Archiving and Processing of Biological Samples for Human Identification 2002 Poster 13th International Symposium on Human ID New advances for FTA technology. PCR, DNA extraction, DNA IQ, hair samples, automation, 96 well FTA format Smuts & Pogue DNA from Urine as a Potential Source of Identification 1999 Poster 10th International Symposium on Human ID Human urine was spotted on FTA cards then amplified by PCR using the Profiler Plus and Cofiler kits. nuclear DNA, mitochondrial DNA, male, female, STR, PCR, FTA Reference DatabaseJun05 29

Snowden et al. Simple, Filter-based PCR Detection of Thelohania solenopsae (Microspora) in Fire Ants (Solenopsis invicta 2002 Journal of Eukaryotic Microbiology, 49(6): 447-448. Application of ant/parasite homogenate to FTA Cards, and comparing that to the conventional methods of extraction. They were looking for a microsporidian parasite in fire ants. 20 ul of ant homogenate on FTA FTA detected fewer spores than silica based DNA purification. spores, microsporidian, parasite, bead beating homogenate, ssu rrna gene, Subrungruang et al. Evaluation of DNA Extraction and PCR Methods for Detection of Enterocytozoon bienuesi in Stool Specimens 2004 J Clin. Microbiol 42(8)3490-3494 Three methods to extract DNA from stool samples were Feces, fecal, parasite, compared; FTA, QIAamp stool mini and chloroform/phenol diagnostic, protozoan extraction. FTA and QIAamp more sensitive than ch/ph. FTA was chosen due to ease, high sensitivity, low cost and less amount of sample required compared to QIAamp kit. Sullivan Comparison Study of DNA extraction and analysis from IsoCode cards, IsoCode Sample Registration Matrix cards and FTA cards and Filter Matrix Bloodstain cards. 2003 MA Thesis California State Univ. Los Angeles, CA Post mortem blood from autopsies applied to FTA, IsoCode, S&S, Whatman BloodStain cards and IsoCode cards. Buccal registration matrix, cells from volunteers applied to IsoCode Sample chelex, extracted DNA, Registration Matrix. DNA was extracted from all media by blood, buccal, postmortem samples chelex. Chelex was significantly better than IsoCode's DNA extraction protocol with 95 o C in water. IsoCode was comparable in number of STR identifed compared to samples on FTA and Bloodstain Cards. Study was done to initially validate IsoCode for the Medical Examiners Office. Taback et al. Prognostic Significance of Circulating Microsatellite Markers in the Plasma of Melanoma Patients. 2001 Cancer Res. 61:5723-5726. FTA used to collect blood from control patients in clinical study. human, diagnostic, cancer study, Taback et al. Detection of Tumor-Specific Genetic Alterations in Bone Marrow from Early-Stage Breast Cancer Patients. 2003 Cancer Res. 63:1884-1887. Whole blood was applied to FTA as a source of control genomic DNA. Portions of genes involved in tumor suppression and metastasis were analyzed from control and tumor derived DNA samples by PCR. human, diagnostic, cancer study, Thacker et al. Use of FTA Cards in Small Volume PCR Reactions 1999 Progress in Forensic Genetics 8, 473-475. One full or a half 1.2mm discs amplified in 5 and 10ml PCR mixes. Compare to chelex extracted DNA. Discs washed in a flat bottom multiwell plate on a shaker using 3 x 100ml FTA Purification Reagent and 2 x 100 ml TE washes. Powerplex 1.2, chelex, AmpFƒSTR Blue, FTA Reference DatabaseJun05 30

Thompson et al. Congenital myasthenic syndrome of Brahman cattle in South Africa Tilsala- Rapid DNA preparation from milk and Timisjarvi and dairy process samples for the Alatossava detection of bacteria by PCR. Tsukaya H A Simple Method for Collecting DNA Samples in the Field 2003 Vet. Rec. 153(4): 779-781. Cattle blood (from EDTA vacutainer) and semen were applied to FTA cards. Punches of 2mm were PCR amplified in 25ml. Semen was diluted 1:10 before application. Punches with semen were resuspended in 200ml FTA Reagent, 20ml 1M dithiothreitol and 5ml 20mg/ml Proteinase K for 1 hr at 56 o C then washed with FTA Reagent and TE-1 as usual before PCR. 2004 Food Microbiol, 21:365-368 Liquid dairy samples or homogenized solid diary samples 2003 Newsletter of Himalayan Botany 32:15-17. were applied as 20-40 ml to FTA. Two 2 mm punches were taken washed and amplified directly by PCR for the detection of probiotic and pathogenic microbes. " the FTA method is easier to perform and...doesn't not include the use of volatile solvents or other toxic reagents." Describes FTA as the most convenient method of collecting plant specimens in the field. Compared FTA to Sigma REDEXtract-N-Amp Plant kits on the basis of room temp stability of DNA and no centrifugation. Leaves from several plant species, common weeds, succulent plants, woody plants and one with a high polysaccharide content (a problem for DNA isolation) were collected with both kits. Author washed the whole card then took a 1 x 2 mm segment for PCR in 50 ul reaction. Efficiency of PCR with FTA was better than Sigma kit but Sigma kit was more precise (less non specific junk bands on gel). The Sigma kit did not work with plants with high polysaccharides or phenolic compounds. FTA worked well with all samples. "FTA Cards are highly recommended for DNA collection in the field...". Breeding cattle, diagnostic, animal biotechnology, gene deletion, screening test bacteria, environmental, milk, cheese, yogurt, clinical mastitis milk samples, dairy st Plant samples, phenol, polysaccharides, Extract- N-Amp Tsukaya H Gene Flow Between Impatients Radicans and I. Javensis (Balsaminaceae) in Gunung Pangrango, Central Java, Indonesia 2004 American Journal of Botany 91(12)2119-2123 FTA cards were used to collect leaf presses from the plants. The whole card was washed and a 1 x 2mm section was taken for PCR amplification in 50ul plant, leaf presses, PCR FTA Reference DatabaseJun05 31

Tsukaya et al Vanek et al. Visapää et al. Large-Scale general collection of wild- 2005 J Plant Res. Online First Project to collect 355 herbarium samples and DNA plant DNA on Mustang, Nepal Czech population data on 10 short tandem repeat loci of SGM Plus STR system kit using DNA Purified in FTA cards Increased cancer risk in heavy drinkers with the alcohol dehydrogenase 1C*1 allele, possibly due to salivary acetaldehyde. 2000 Forensic Science International 119:107-108 specimens from Nepal to be included in the Flora of Nepal Database website. Plant leaves were pressed onto FTA, washed and 1mm 2 sections were amplified by PCR. The authors drew a grid on a classic card to store 20 samples/card and found no cross contamination. The Flora data base may be using FTA to send DNA samples out to researchers. Population study on 10 STR loci on 202 unrelated Czech Caucasians. 2004 Gut 53:871-876. FTA was used to collect whole blood samples from patients with cancer and normal control patients. Three 2 mm disks were washed with FTA reagent then TE-1 followed by drying at 60oC for 30 min. Portions of the alcohol dehydrogenase gene were amplified by PCR then digested for RFLP analysis. plant, collections, PCR, DNA database Population study, blood, human ID population study, blood, human ID, allele frequency, cancer predisposition Waiswa et al. Monitor lizard (Varanus niloticus, Linnaeus, 1766) as a host for tsetse (Glossina fuscipes fuscipes, Newstead, 1910) in the sleeping sickness endemic foci of Uganda. 2003 African Journal of Ecology 41,349-351. Reference blood samples from monitor lizards and tsetse smears collected onto FTA. This publication's main analysis was ELISA to discriminate tsetse species. No DNA work was presented. Wang et al. SNP Analysis Based on Strand Displacement Amplification 2004 IVD Technol. 10(6)61-71 Work from BD Diagnostics describing BD ProbeTec ET system that looks at SNPs via a whole genome amplification approach instead of other SNP methods like primer extension or microarray. Blood on FTA serves as a good DNA template for this platform. Paper can be accessed at www.devicelink.com/ivdt/archive/04/07/012.html diagnostics, WGA, Williams et al. Automation of in situ Sample Preparation for PCR Using the FTA DNA Collection System and the Rosys Laboratory Workstation 1996 Poster Advances in Forensic Haemogenetics 6 Illustrates the Rosys GeneMachine for the processing of DNA on FTA cards for STR profiling. FTA superior to 903. automation, liquid handling, blood, STR, 903 v FTA FTA Reference DatabaseJun05 32

Yet et al. Comparison of DNA Recovered from FTA Paper to Conventional Organic Extraction Procedures using a Hae III Based RFLP System. 1997 Poster: 8th Annual Int. Symposium of Human Identification DNA collected and archived on FTA performed well for RFLP analysis. RFLP, archiving, preservation Zhong et al. Comparison of IsoCode STX and FTA Gene Guard Collection Matrices as Whole Blood Storage and Processing Devices for the Diagnosis of Malaria by PCR 2001 Journal of Clinical Microbiology 39: 1195-1196 FTA outperforms IsoCode by >20% for the detection of multiple species malaria 96% detection of single species malaria. diagnosis, transportation, parasites, plasmodial small subunit RNA gene, Plasmodium vivax, single species infections, mixed infections Zhou and Hickford Zhou H, Hickford JGH and Fang Q Allelic Polymorphism in the Ovine DQA1 gene Technical Note: Dtermination of alleles of the ovine PRNP gene using PCR- single-strand conformational polymorphism analysis. 2004 J. Anim Sci. 82:8-16 Blood from 300 sheep was collected using FTA cards. The gene for DQA1 was amplified by PCR and sequenced. 2005 J. Anim. Sci 83:745-749. Blood from 400 sheep covering 6 breeds was collected onto FTA cards to amplify a 173bp fragment of the prion gene. Codons 136, 154 and 171 are tested to determine an animal's susceptibility to scrapie. Punches of 1.2mm were included in the 20µl PCR mix. The selected amplicons were inserted into plasmids and sequenced. genetic analysis, agriculture, animal, sheep, SSCP, rapid genotyping method, prion protein gene Zhou H, Hickford JGH and Fang Q Polymorphism of the DQA2 gene in goats 2005 J. Anim Sci 83:963-968. Blood samples were collected from Boer goats onto FTA. Punches, 1.2mm, were washed according to the standard protocol and included in 20µl PCR amplifications. caprine, goats, single stranded conformational polymorphism, SSCP, cloning, sequencing FTA Reference DatabaseJun05 33

FTA Blood DNA Whatman Raccolta, trasferimento, stoccaggio e purificazione di DNA da campioni di sangue FTA della Whatman offre un valido e semplice metodo per la preparazione di campioni per analisi di DNA da sangue. Occorre, infatti, semplicemente applicare un campione di sangue, midollo osseo o buffy coat alla matrice FTA. Il DNA viene catturato e stabilizzato all istante, consentendone lo stoccaggio per un periodo indefinito a temperatura ambiente, da analizzare in qualsiasi momento. Trasporto sicuro e purificazione in 30 minuti rendono la tecnologia FTA un indispensabile strumento di ricerca. Caratteristiche Principali Semplice raccolta dei campioni I campioni vengono depositati direttamente sulla matrice FTA. Purificazione rapida FTA rappresenta il metodo più veloce per la purificazione di DNA da campioni di sangue per PCR. Il DNA viene purificato sulla FTA Card in tre semplici steps, in un singolo tubo, a temperatura ambiente. Non sono necessarie sostanze chimiche tossiche, inoltre rimanendo il DNA fissato sulla matrice è subito pronto per la PCR. Adatte a qualsiasi metodo Le FTA Cards si adattano alle diverse tipologie di campione: preparazione di campioni di sangue, midollo osseo o buffy coat. E sufficiente applicare direttamente i campioni sulla matrice FTA e non è necessaria alcuna ulteriore purificazione nè con sostanze tossiche (metodi quali fenolo/cloroformio) nè con altri metodi che possono richiedere lunghe incubazioni. Applicazioni Applicazioni diagnostiche e cliniche Tipizzazione tissutale HLA Identificazione genetica Stoccaggio a temperatura ambiente e trasferimento sicuri Gli acidi nucleici vengono automaticamente stabilizzati senza la necessità di congelarli. FTA inattiva gli agenti patogeni potenzialmente nocivi rendendo i campioni sicuri per la loro manipolazione in laboratorio. Inviare i campioni a colleghi o al laboratorio centrale è veramente facile con FTA Cards. Occorre solo inviarli per posta! Automatizzabile L utilizzo di sistemi automatici accelera le fasi di preparazione di diversi dischi prelevati contemporaneamente, ottimizza le risorse disponibili e standardizza la purificazione di DNA. I dischi prelevati dalla FTA Card possono essere sottoposti alle fasi di lavaggio e preparazione per la PCR su una diversi tipi di strumenti presenti in commercio (liquid handling robot). Studi di screening molecolari Studi di allevamento Whole genomic amplification TRE SEMPLICI PASSAGGI OPERATIVI PER OTTENERE UN DNA PURO Purificare Dispensare il campione Prelevare un piccolo disco (punch) dal Campione

PROTOCOLLO APPLICATIVO DEL FTA BLOOD DNA INFORMAZIONI PER L ORDINE DAL CATALOGO WHATMAN No. Cat. WB120061 WB120204 WB120205 WB120055 WB120210 WB120208 WB100005 WB100006 WB100028 WB100010 WB100011 WB120216 WB120217 WB100025 WB100003 WB100016 WB100020 WB100030 WB120005 Dispensare campione Dispensare il campione di sangue, midollo osseo o buffy coat sulla FTA Card. Lasciarlo asciugare completamente. Prelevare un piccolo disco Prelevare (mediante perforazione) un piccolo disco dal campione depositato sulla FTA Card. Purificazione con Reagente di Purificazione FTA (Purification Reagent FTA) Mettere il disco di campione in un tubo per PCR ed effettuare tre lavaggi con il Reagente di Purificazione FTA. Eliminare la soluzione utilizzata dopo ciascun lavaggio. Whatman GenSpin Kit di Purificazione DNA Descrizione FTA Starter Pack Reagente di Purificazione FTA FTA Classic Card (non-indicatrice) FTA Mini Card (non-indicatrice) FTA Micro Card (non-indicatrice) FTA Gene Card Harris Micro Perforatore 1.2mm Harris Micro Perforatore 1.2mm Tip Harris Uni-Core 1.25mm Perforatore Busta Multi-Barrier (grande) Busta Multi-Barrier (piccola) FTA Gene Card/Tasca/Disseccante FTA Classic Card/Tasca/Disseccante 1.2mm Plunger di Ricambio Essiccante (1g) Busta Postale per FTA Card Tagliere di Ricambio Supporto per FTA Gene Card GenSpin Kit di Purificazione DNA Lavaggio con soluzione tampone TE-1 Effettuare due lavaggi con la soluzione tampone TE-1 (10mM Tris, 0,1 mm EDTA, ph 8,0). Eliminare la soluzione usata dopo ciascun lavaggio. Essiccazione Lasciar asciugare il disco nel tubo PCR PCR Aggiungere la master mix per la PCR direttamente al disco e iniziare la reazione di amplificazione. Quantità per Confezione 1 500mL 100 100 100 100 1 1 4 500 500 1000 1000 1 1000 50 1 20 50 Purificazioni Purificazione del DNA in meno di 25 minuti! Il GenSpin Kit di Purificazione del DNA della Whatman rappresenta un metodo estremamente semplice per purificare il DNA a singolo filamento da campioni di sangue e cellule in coltura. Il campione è subito pronto per la PCR a partire anche da piccoli volumi del campione di partenza. Qualità Whatman Whatman, azienda leader nelle teconologie di seprazione, è nota nel settore scientifico per prodotti e soluzioni innovative contraddistinte da un elevata qualità. La certezza del valore di una sempre maggior semplificazione delle procedure di laboratorio tende ad accellerare lo sviluppo di nuovi prodotti, a ridurre i costi e tempi di processo. Inoltre, per accentuare ulteriormente l impegno nel soddisfare esigenze specifiche dei consumatori, Whatman è organizzata in quattro aree distinte: Chimica Analitica, Diagnostica, Genomics e Proteomica, e Dispositivi Medicali. Per ulteriori informazioni vogliate collegarvi al sito www.whatman.com. FTA, GenSpin e Whatman sono marchi registrati della Whatman Group. North America Europe Japan Asia Pacific Whatman Inc 9 Bridewell Place Clifton, NJ 07014 USA Technical Support: 1-800-922-0361 Customer Service: 1-800-631-7290 Outside US: 973-773-5800 Fax: 973-773-0168 E-mail: info@whatman.com Whatman International Ltd Springfield Mill James Whatman Way Maidstone Kent ME14 2LE UK Tel: +44 (0)1622 676670 Fax: +44 (0)1622 677011 E-mail: information@whatman.com Whatman Japan KK Daiwa Ueno Building 1F 6-10 Ueno 5-chome, Taito-ku Tokyo 110-0005, Japan Tel: +81 (0)3 3832 6707 Fax: +81 (0)3 3832 6457 E-mail: japaninfo@whatman.com Whatman Asia Pacific Pte Ltd 171 Chin Swee Road #08-01 San Centre Singapore 169877 Tel: +65 6534 0138 Fax: +65 6534 2166 E-mail: wap@whatman.com Leaders in Separations Technology www.whatman.com Cat No. S9036-779

FTA Total RNA Whatman Raccolta, trasferimento, stoccaggio e purificazione di RNA Totale FTA della Whatman offre un valido e semplice metodo per la preparazione di RNA Totale da una varietà di tipologia di campioni di partenza per analisi molecolari. Occorre, infatti, semplicemente applicare un campione di sangue, di cellule in coltura, o pressare foglie sulla matrice FTA. Gli acidi nucleici sono catturati e stabilizzati all istante, mentre agenti patogeni e nucleasi vengono inattivati. Il trasferimento e lo stoccaggio dei campioni sono sicuri fino al momento dell analisi. Purificazione di RNA per RT-PCR risulta estremamente semplificata. Provate FTA e lo troverete immediatamente un indispensabile strumento per il laboratorio di Biologia Molecolare. Caratteristiche Principali Tre semplici passaggi operativi per RNA Totale RNA adatto per Transcriptasi- Inversa (RT)-PCR e Northern Blotting RNA purificato da FTA può essere utilizzato come stampo nelle analisi RT-PCR direttamente in soluzione, oppure può essere precipitato e caricato su gel di agarosio per analisi di Northern Blotting. Raccolta I campioni sono applicati direttamente sulla matrice FTA. Gli acidi nucleici vengono automaticamente stabilizzati e protetti: per un breve stoccaggio a temperatura ambiente e, per periodi più estesi di stoccaggio, a temperature di -20ºC o -70ºC. Stoccaggio a temperatura ambiente e trasferimento sicuri FTA inattiva agenti patogeni nocivi rendendo i campioni sicuri per la manipolazione in laboratorio e per il trasferimento. Rapida purificazione RNA viene eluito interamente dalla FTA Card mediante un semplice step di lavaggio e dispensato in un singolo tubo, a temperatura ambiente. Non richiede l utilizzo di sostanze chimiche tossiche. RNA viene eluito dalla matrice ed è subito pronto per reazioni di RT-PCR. Adatto al vostro metodo Non richiede ulteriori procedure di purificazione con lunghi metodi quali fenolo/cloroformio, fasi di digestione o disgregazione di tessuti. Dispensare il Campione Purificare Prelevare mediante perforazione un piccolo disco (punch) dal Campione Applicazioni Applicazioni di ricerca RT/PCR Northern Blotting Espressione genica PCR real time

PROTOCOLLO APPLICATIVO DEL FTA TOTAL RNA Dispensare Campione Dispensare il campione sulla FTA Card. Lasciarlo asciugare completamente. Eluizione del RNA con Soluzione Tampone Aggiungere la soluzione tampone, incubare in ghiaccio. Prelevare un piccolo disco Prelevare mediante perforazione un piccolo disco (punch) dal Campione sulla FTA Card e inserirlo in un tubo da 1,5mL Perforare estraendo un disco dal campione sulla FTA Card e inserirlo in un tubo da 1,5mL. RT-PCR o Northern Blotting Usare RNA direttamente per RT-PCR oppure precipitare per analisi Nothern Blot. RT-PCR da FTA: RNA da cellule HL60 estratte con FTA o con metodo Competitor T MW 1 2 3 4 5 6 Qualità Whatman Whatman, azienda leader nelle tecnologie per la separazione, è nota nellà comunità scientifica per prodotti e soluzioni innovative contraddistinte da un elevata qualità. Il nostro istinto per la semplificazione accelera la velocità di fare nuove scoperte, riduce i costi e risparmia tempo. Inoltre, per accentuare ulteriormente l impegno nel soddisfare le specifiche esigenze dei nostri clienti, Whatman è organizzata in quattro aree distinte: LabScience, BioScience, MedTech Diagnostica e MedTech Dispoitivi. Ulteriori informazioni sono disponibili collegandovi al sito www.whatman.com. FTA, GenSpin e Whatman sono marchi registrati della Whatman Group. INFORMAZIONI PER L ORDINE DAL CATALOGO WHATMAN No. Cat. Linea 1 & 2: PCR utilizzando RNA da FTA (nessuna contaminazione da DNA). Linea 3: Controllo Negativo (nessuno stampo). Linea 4: PCR utilizzando RNA da Competitor T (contaminazione DNA). Linea 5: PCR utilizzando DNA controllo positivo K562. Linea 6: RT-PCR utilizzando RNA da FTA. Descrizione Quantità per Confezione WB120061 FTA Starter Pack 1 WB120205 FTA Classic Card (non-indicatrice) 100 WB120206 FTA Classic Card (indicatrice) 100 WB120055 FTA Mini Card (non-indicatrice) 100 WB120056 FTA Mini Card (indicatrice) 100 WB120210 FTA Micro Card (non-indicatrice) 100 WB120211 FTA Micro Card (indicatrice) 100 WB120208 FTA Gene Card 100 WB100007 Harris Micro Perforatore 2.0mm 1 WB100008 Harris Micro Perforatore 2.0mm Tip 1 WB100029 Harris Uni-Core 2.0mm Perforatore 4 WB100010 Busta Multi-Barrier (grande) 500 WB100011 Busta Multi-Barrier (piccola) 500 WB120216 FTA Gene Card/Busta/Essiccante 1000 WB120217 FTA Classic Card/Busta/Essiccante 1000 WB100026 2.0mm Plunger di Ricambio 1 WB100003 Essiccante (1g) 1000 WB100016 Busta Postale per FTA Card 50 WB100020 Tagliere di Ricambio 1 WB100030 Supporto per FTA Gene Card 20 North America Europe Japan Asia Pacific Whatman Inc 9 Bridewell Place, Clifton, NJ 07014 Technical Support: 1-800-922-0361 Customer Service: 1-800-631-7290 Outside US: 973-773-5800 Fax: 973-773-0168 E-mail: info@whatman.com Whatman International Ltd Springfield Mill James Whatman Way Maidstone Kent ME14 2LE, UK Tel: +44 (0)1622 676670 Fax: +44 (0)1622 691425 E-mail: information@whatman.com Whatman Japan KK Daiwa Ueno Building 1F 6-10 Ueno 5-chome, Taito-ku Tokyo 110-0005, Japan Tel: +81 (0)3 3832 6707 Fax: +81 (0)3 3832 6457 E-mail: japaninfo@whatman.com Whatman Asia Pacific Pte Ltd 171 Chin Swee Road #08-01 San Centre Singapore 169877 Tel: +65 6534 0138 Fax: +65 6534 2166 E-mail: wap@whatman.com Leaders in Separations Technology www.whatman.com Cat No. S9036-785