BioBank TM cdna Kit. Instructions for the use of BioBank TM cdna in real-time PCR. BB BioBank control cdna

Save this PDF as:
 WORD  PNG  TXT  JPG

Size: px
Start display at page:

Download "BioBank TM cdna Kit. Instructions for the use of BioBank TM cdna in real-time PCR. BB BioBank control cdna"

Transcription

1 BioBank TM cdna Kit Instructions for the use of BioBank TM cdna in real-time PCR BB BioBank control cdna

2 Contents Introduction 3 Kit Contents 4 Reagents and Equipment to Be Supplied by User 4 PrimerDesign Satisfaction Guarantee 5 Quality Control 5 Bench-side Protocol 6 Amplification Protocol 8 2

3 Introduction The BioBank TM a high quality source of cdna validated for use in real-time PCR experiments. The cdna is reverse transcribed from high quality DNAse treated RNA from a variety of sources using an optimised blend of Oligo-dT and random nonamer primers. BioBank TM cdna is therefore essentially free of genomic DNA and PCR inhibitors and covers the widest possible range of RNA and mrna transcripts in the specified tissue or cell line. BioBank TM cdna is useful for expression profiling of newly identified genes, and also as a positive control for the real-time PCR step. Positive control Primers Positive control primers and probe are supplied with the kits which detect 18S ribosomal RNA. These primers include a Double dye probe for optimal probe based detection through the FAM channel. The positive control primer and probe mix can be used with standard Taqman cycling conditions and with SYBRgreen mastermix for detection using SYBR green chemistry. The positive control primers will give a cycle threshold value of approximately CT=12 depending on the detection chemistry and the particular hardware settings in use. 3

4 Kit Contents BioBank TM cdna 250ng (PURPLE) Control primers and probe for 18S ribosomal RNA (BROWN) RNAse/DNAse free water Reagents and Equipment to Be Supplied by User Real-Time PCR instrument Mastermix or Mastermix components This kit is designed to work well with all commercially available Mastermixes. However, we recommend the use of PrimerDesign 2x PrecisionPLUS TM TM or PrecisionFAS Mastermix. Pipettors and Tips Vortex and centrifuge Kit Storage The PrimerDesign BioBank TM cdna kit should be stored at -20ºC on arrival. Freeze/thawing cycles should be kept to a minimum once resuspended. Under these conditions reagents are stable for six months from date of resuspension. 4

5 PrimerDesign Satisfaction Guarantee PrimerDesign takes pride in the quality of all of our products. Should this product fail to perform satisfactorily when used according to the protocols in this manual, PrimerDesign will replace the item free of charge. Quality Control As part of our ISO9001 and ISO13485 quality assurance systems, all PrimerDesign products are monitored to ensure the highest levels of performance and reliability. 5

6 Bench-side Protocol 1. Pulse-spin each tube in a centrifuge before opening. This will ensure lyophilised primer and probe mix is in the base of the tube and is not spilt upon opening the tube. 2. Resuspend lyophilised primer and probe mix and cdna in RNAse/DNAse free water provided according to the table below; To ensure complete resuspension, vortex each tube thoroughly, allow to stand for 5 minutes and vortex again before use. Component Volume BioBank TM cdna (PURPLE) 130 µl 18S control primers and probe (BROWN) 50 µl Real-time PCR 1. Positive control with 18S Make up a well in duplicate containing the positive control primers and BioBank TM cdna Component 1 Reaction PrecisionPLUS TM TM or PrecisionFAS Mastermix 10 µl 18S primers and probe (BROWN) 1 µl BioBank TM cdna (PURPLE) 5 µl RNAse/DNAse free water (WHITE) 4 µl Final volume 20 µl 2. Target gene with PrimerDesign custom designed real-time PCR assay (optional step) Make up a well in duplicate containing the positive control primers and BioBank TM cdna Component 1 Reaction PrecisionPLUS TM TM or PrecisionFAS Mastermix 10 µl Target gene primer mix 1 µl BioBank TM cdna (PURPLE) 5 µl RNAse/DNAse free water (WHITE) 4 µl Final volume 20 µl 6

7 3. Target gene with experimental primers (optional step) Make up a well in duplicate containing the experimental primers and BioBank TM cdna Component 1 Reaction PrecisionPLUS TM TM or PrecisionFAS Mastermix 10 µl Forward primer 6pmol Reverse primer 6pmol Probe (optional) 3pmol BioBank TM cdna (PURPLE) 5 µl RNAse/DNAse free water (WHITE) X µl Final volume 20 µl 7

8 Amplification Protocol Amplification conditions using Taqman or SYBR cycling settings with PrecisionPLUS TM mastermix Cycling x40* Step Time Temp Enzyme activation 2min 95ºC Denaturation 15s 95ºC DATA COLLECTION** 60s 60ºC * For low copy number targets, giving late detection, a further 10 cycles may be needed to generate the complete amplification plot. ** Fluorogenic data should be collect during this step through the FAM channel. Amplification conditions using Taqman or SYBR cycling settings with PrecisionFAS TM mastermix Cycling x40* Step Time Temp Enzyme activation 2min 95ºC Denaturation 5s 95ºC DATA COLLECTION** 20s 60ºC * For low copy number targets, giving late detection, a further 10 cycles may be needed to generate the complete amplification plot. ** Fluorogenic data should be collect during this step through the FAM channel. 8

Genomic DNA detection assay

Genomic DNA detection assay Genomic DNA detection assay Detection of genomic DNA by real-time PCR Contents CTRL Internal controls and gdna detection Contents Kit Contents 3 Reagents and Equipment to Be Supplied by User 3 Kit Storage

More information

oasig TM lyophilised 2X qpcr Mastermix

oasig TM lyophilised 2X qpcr Mastermix oasig TM lyophilised 2X qpcr Mastermix Instructions for use of Primerdesign oasig lyophilised Mastermix MM MasterMix Contents Introduction 3 Kit Contents 4 Kit Storage 4 Suitable Sample Material 4 Licensing

More information

Precision TM FAST and Precision TM Mastermix

Precision TM FAST and Precision TM Mastermix Precision TM FAST and Precision TM Mastermix Instructions for use of PrimerDesign Precision TM Mastermix and Precision TM FAST Mastermix MM MasterMix Contents Introduction 3 Kit Contents 5 Recommended

More information

Precision OneStep TM qrt-pcr Mastermix

Precision OneStep TM qrt-pcr Mastermix Precision OneStep TM qrt-pcr Mastermix Instructions for use of Primerdesign Precision OneStep TM Mastermix for real-time RT-PCR ONE OneStep MasterMix Contents Introduction 3 Kit Contents 5 Recommended

More information

PrecisionPLUS TM PrecisionFAST TM Precision TM Mastermix

PrecisionPLUS TM PrecisionFAST TM Precision TM Mastermix PrecisionPLUS TM PrecisionFAST TM Precision TM Mastermix Instructions for use of PrimerDesign Precision TM Mastermix/Precision Plus TM Precision TM FAST Mastermix Mastermix and MM MasterMix Contents Introduction

More information

Dengue Virus subtypes 1,2 3 and 4. genesig Standard Kit. DNA testing. Everything... Everyone... Everywhere... 3 Untranslated Region (3 UTR) 150 tests

Dengue Virus subtypes 1,2 3 and 4. genesig Standard Kit. DNA testing. Everything... Everyone... Everywhere... 3 Untranslated Region (3 UTR) 150 tests TM Primerdesign Ltd TM Primerdesign Ltd Dengue Virus subtypes 1,2 3 and 4 3 Untranslated Region (3 UTR) genesig Standard Kit 150 tests DNA testing Everything... Everyone... Everywhere... For general laboratory

More information

Epstein Barr Virus (Human Herpes virus 4) genesig Standard Kit. DNA testing. Everything... Everyone... Everywhere...

Epstein Barr Virus (Human Herpes virus 4) genesig Standard Kit. DNA testing. Everything... Everyone... Everywhere... TM Primerdesign Ltd TM Primerdesign Ltd Epstein Barr Virus (Human Herpes virus 4) nonglycosylated membrane protein (BNRF1) gene genesig Standard Kit 150 tests DNA testing Everything... Everyone... Everywhere...

More information

Human Herpes Virus 1 (Herpes simplex type 1) genesig Standard Kit. DNA testing. Everything... Everyone... Everywhere...

Human Herpes Virus 1 (Herpes simplex type 1) genesig Standard Kit. DNA testing. Everything... Everyone... Everywhere... TM Primerdesign Ltd TM Primerdesign Ltd Human Herpes Virus 1 (Herpes simplex type 1) Capsid assembly and DNA maturation gene genesig Standard Kit 150 tests DNA testing Everything... Everyone... Everywhere...

More information

Hepatitis B Virus. genesig Advanced Kit. DNA testing. Everything... Everyone... Everywhere... Core Protein Region. 150 tests.

Hepatitis B Virus. genesig Advanced Kit. DNA testing. Everything... Everyone... Everywhere... Core Protein Region. 150 tests. TM Primerdesign Ltd TM Primerdesign Ltd Hepatitis B Virus Core Protein Region genesig Advanced Kit 150 tests DNA testing Everything... Everyone... Everywhere... For general laboratory and research use

More information

Epstein Barr Virus (Human Herpes virus 4) nonglycosylated membrane protein (BNRF1) gene. genesig Advanced Kit. DNA testing

Epstein Barr Virus (Human Herpes virus 4) nonglycosylated membrane protein (BNRF1) gene. genesig Advanced Kit. DNA testing TM Primerdesign Ltd TM Primerdesign Ltd Epstein Barr Virus (Human Herpes virus 4) nonglycosylated membrane protein (BNRF1) gene genesig Advanced Kit 150 tests DNA testing Everything... Everyone... Everywhere...

More information

Lyme Disease. genesig Standard Kit. DNA testing. Everything... Everyone... Everywhere... RecA gene. 150 tests. Primerdesign Ltd.

Lyme Disease. genesig Standard Kit. DNA testing. Everything... Everyone... Everywhere... RecA gene. 150 tests. Primerdesign Ltd. TM Primerdesign Ltd TM Primerdesign Ltd Lyme Disease RecA gene genesig Standard Kit 150 tests DNA testing Everything... Everyone... Everywhere... For general laboratory and research use only 1 Introduction

More information

Klebsiella oxytoca. genesig Standard Kit. DNA testing. Everything... Everyone... Everywhere... Outer membrane protein II (ompa) gene.

Klebsiella oxytoca. genesig Standard Kit. DNA testing. Everything... Everyone... Everywhere... Outer membrane protein II (ompa) gene. TM Primerdesign Ltd TM Primerdesign Ltd Klebsiella oxytoca Outer membrane protein II (ompa) gene genesig Standard Kit 150 tests DNA testing Everything... Everyone... Everywhere... For general laboratory

More information

Quantification of Propionibacterium acnes. Standard kit 150 tests

Quantification of Propionibacterium acnes. Standard kit 150 tests Primerdesign TM Ltd Quantification of Propionibacterium acnes Recombinase A (reca) gene For general laboratory and research use only Standard kit 150 tests Introduction to Propionibacterium acnes Propionibacterium

More information

Human Herpes Virus 4 (Epstein Barr)

Human Herpes Virus 4 (Epstein Barr) Techne qpcr test Human Herpes Virus 4 (Epstein Barr) nonglycosylated membrane protein (BNRF1) gene 150 tests For general laboratory and research use only 1 Introduction to Human Herpes Virus 4 (Epstein

More information

Real-time quantitative RT -PCR (Taqman)

Real-time quantitative RT -PCR (Taqman) Real-time quantitative RT -PCR (Taqman) Author: SC, Patti Lab, 3/03 This is performed as a 2-step reaction: 1. cdna synthesis from DNase 1-treated total RNA 2. PCR 1. cdna synthesis (Advantage RT-for-PCR

More information

Lyme Disease RecA gene. genesig Advanced Kit. DNA testing. Everything... Everyone... Everywhere... 150 tests. Primerdesign Ltd.

Lyme Disease RecA gene. genesig Advanced Kit. DNA testing. Everything... Everyone... Everywhere... 150 tests. Primerdesign Ltd. TM Primerdesign Ltd TM Primerdesign Ltd Lyme Disease RecA gene genesig Advanced Kit 150 tests DNA testing Everything... Everyone... Everywhere... For general laboratory and research use only 1 Introduction

More information

Lyme Disease. RecA gene. 150 tests. Quantification of Lyme Disease genomes. Advanced kit handbook HB10.03.07

Lyme Disease. RecA gene. 150 tests. Quantification of Lyme Disease genomes. Advanced kit handbook HB10.03.07 Techne qpcr test Lyme Disease RecA gene 150 tests For general laboratory and research use only 1 Introduction to Lyme Disease Lyme disease is an infectious disease caused mainly by three species of bacteria

More information

Quantification of Hepatitis B Virus. Core Protein Region. 150 tests

Quantification of Hepatitis B Virus. Core Protein Region. 150 tests Quantification of Hepatitis B Virus Core Protein Region For general laboratory and research use only 150 tests Introduction to Hepatitis B Virus Originally known as serum hepatitis, hepatitis B has only

More information

Quantification of Mycobacterium Tuberculosis. 150 tests

Quantification of Mycobacterium Tuberculosis. 150 tests Quantification of Mycobacterium Tuberculosis rep 13E12 For general laboratory and research use only 150 tests Introduction to Mycobacterium Tuberculosis Mycobacterium tuberculosis is the bacterium that

More information

Borrelia burgdorferi

Borrelia burgdorferi Techne qpcr test Borrelia burgdorferi Outer surface protein A (ospa) gene 150 tests For general laboratory and research use only 1 Introduction to Borrelia burgdorferi Borrelia burgdorferi is a Gram negative

More information

First Strand cdna Synthesis

First Strand cdna Synthesis 380PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name First Strand cdna Synthesis (Cat. # 786 812) think proteins! think G-Biosciences

More information

Streptococcus pneumoniae

Streptococcus pneumoniae Techne qpcr test Streptococcus pneumoniae alpha-fucosidase gene 150 tests For general laboratory and research use only 1 Introduction to Streptococcus pneumoniae Streptococcus pneumoniae, or pneumococcus,

More information

Herpes Simplex Virus 1 and 2

Herpes Simplex Virus 1 and 2 Techne qpcr test Herpes Simplex Virus 1 and 2 HSV1 - UL23 thymidine kinase (TK) gene HSV2 - UL36 large tegument protein gene 150 tests For general laboratory and research use only 1 Introduction to Herpes

More information

Klebsiella oxytoca. Outer membrane protein II (ompa) gene. 150 tests. Quantification of Klebsiella oxytoca genomes. Advanced kit handbook HB10.03.

Klebsiella oxytoca. Outer membrane protein II (ompa) gene. 150 tests. Quantification of Klebsiella oxytoca genomes. Advanced kit handbook HB10.03. Techne qpcr test Klebsiella oxytoca Outer membrane protein II (ompa) gene 150 tests For general laboratory and research use only 1 Introduction to Klebsiella oxytoca Klebsiella oxytoca is a Gram-negative,

More information

Thermo Scientific DyNAmo cdna Synthesis Kit for qrt-pcr Technical Manual

Thermo Scientific DyNAmo cdna Synthesis Kit for qrt-pcr Technical Manual Thermo Scientific DyNAmo cdna Synthesis Kit for qrt-pcr Technical Manual F- 470S 20 cdna synthesis reactions (20 µl each) F- 470L 100 cdna synthesis reactions (20 µl each) Table of contents 1. Description...

More information

PrimeScript RT Master Mix (Perfect Real Time) Sample

PrimeScript RT Master Mix (Perfect Real Time) Sample Cat. # RR036Q For Research Use PrimeScript RT Master Mix (Perfect Real Time) Sample Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Materials Required but not Provided...

More information

Considerations for standardizing qpcr assays.

Considerations for standardizing qpcr assays. Considerations for standardizing qpcr assays. qpcr Satellite Symposium BioCity Leipzig March 10-11, 11, 2005 Reinhold Mueller, PhD Senior Staff Scientist Outline: Introduction Controls, references and

More information

Introduction To Real Time Quantitative PCR (qpcr)

Introduction To Real Time Quantitative PCR (qpcr) Introduction To Real Time Quantitative PCR (qpcr) SABiosciences, A QIAGEN Company www.sabiosciences.com The Seminar Topics The advantages of qpcr versus conventional PCR Work flow & applications Factors

More information

Super One-step RT-PCR Kit for Virus Detection

Super One-step RT-PCR Kit for Virus Detection Super One-step RT-PCR Kit for Virus Detection For fast and sensitive one-step RT-PCR with highly specificity www.tiangen.com/en SD121221 Super One-step RT-PCR Kit Kit Contents Contents Cat. no. SD103 SD103

More information

Realtime PCR Master Mix

Realtime PCR Master Mix Instruction manual Realtime PCR Master Mix 0810 F0923K Realtime PCR Master Mix Contents [1] Introduction [2] Components [3] Primer/Probe design [4] Detection [5] Specimens [6] Protocol 1. TaqMan assay

More information

Quantitative Real Time PCR Protocol. Stack Lab

Quantitative Real Time PCR Protocol. Stack Lab Quantitative Real Time PCR Protocol Stack Lab Overview Real-time quantitative polymerase chain reaction (qpcr) differs from regular PCR by including in the reaction fluorescent reporter molecules that

More information

Mir-X mirna First-Strand Synthesis Kit User Manual

Mir-X mirna First-Strand Synthesis Kit User Manual User Manual Mir-X mirna First-Strand Synthesis Kit User Manual United States/Canada 800.662.2566 Asia Pacific +1.650.919.7300 Europe +33.(0)1.3904.6880 Japan +81.(0)77.543.6116 Clontech Laboratories, Inc.

More information

All-in-One First-Strand cdna Synthesis Kit

All-in-One First-Strand cdna Synthesis Kit All-in-One First-Strand cdna Synthesis Kit For reliable first-strand cdna synthesis from all RNA sources Cat. No. AORT-0020 (20 synthesis reactions) Cat. No. AORT-0050 (50 synthesis reactions) User Manual

More information

Stratagene QPCR Mouse Reference Total RNA

Stratagene QPCR Mouse Reference Total RNA Stratagene QPCR Mouse Reference Total RNA Instruction Manual Catalog #750600 Revision C.0 For Research Use Only. Not for use in diagnostic procedures. 750600-12 LIMITED PRODUCT WARRANTY This warranty limits

More information

Validating Microarray Data Using RT 2 Real-Time PCR Products

Validating Microarray Data Using RT 2 Real-Time PCR Products Validating Microarray Data Using RT 2 Real-Time PCR Products Introduction: Real-time PCR monitors the amount of amplicon as the reaction occurs. Usually, the amount of product is directly related to the

More information

SYBR Premix Ex Taq (Tli RNaseH Plus)

SYBR Premix Ex Taq (Tli RNaseH Plus) Cat. # RR420A For Research Use SYBR Premix Ex Taq (Tli RNaseH Plus) Product Manual Table of Contents I. Description... 3 II. Principle... 3 III. Components... 4 IV. Materials Required but not Provided...

More information

Reference Dye N/A ROX & ROX II (as separate tubes) ROX & ROX II (as separate tubes)

Reference Dye N/A ROX & ROX II (as separate tubes) ROX & ROX II (as separate tubes) Comparison of TaKaRa s and qrt-pcr Kits SYBR Premix Ex Taq (Tli RNase H+) Premix Ex Taq (Probe qpcr) SYBR Premix Ex Taq II (Tli RNase H+) Cat.# RR420A RR390A RR820A Reactions/Kit 200 (50 µl reaction size)

More information

ab185916 Hi-Fi cdna Synthesis Kit

ab185916 Hi-Fi cdna Synthesis Kit ab185916 Hi-Fi cdna Synthesis Kit Instructions for Use For cdna synthesis from various RNA samples This product is for research use only and is not intended for diagnostic use. Version 1 Last Updated 1

More information

User Manual. CelluLyser Lysis and cdna Synthesis Kit. Version 1.4 Oct 2012 From cells to cdna in one tube

User Manual. CelluLyser Lysis and cdna Synthesis Kit. Version 1.4 Oct 2012 From cells to cdna in one tube User Manual CelluLyser Lysis and cdna Synthesis Kit Version 1.4 Oct 2012 From cells to cdna in one tube CelluLyser Lysis and cdna Synthesis Kit Table of contents Introduction 4 Contents 5 Storage 5 Additionally

More information

DyNAmo cdna Synthesis Kit for qrt-pcr

DyNAmo cdna Synthesis Kit for qrt-pcr DyNAmo cdna Synthesis Kit for qrt-pcr Instruction manual F- 470S Sufficient for 20 cdna synthesis reactions (20 µl each) F- 470L Sufficient for 100 cdna synthesis reactions (20 µl each) Description...

More information

SYBR Green Realtime PCR Master Mix -Plus-

SYBR Green Realtime PCR Master Mix -Plus- Instruction manual SYBR Green Realtime PCR Master Mix -Plus- 0810 F0925K SYBR Green Realtime PCR Master Mix -Plus- Contents QPK-212T 1mLx1 QPK-212 1mLx5 Store at -20 C, protected from light [1] Introduction

More information

REAL TIME PCR USING SYBR GREEN

REAL TIME PCR USING SYBR GREEN REAL TIME PCR USING SYBR GREEN 1 THE PROBLEM NEED TO QUANTITATE DIFFERENCES IN mrna EXPRESSION SMALL AMOUNTS OF mrna LASER CAPTURE SMALL AMOUNTS OF TISSUE PRIMARY CELLS PRECIOUS REAGENTS 2 THE PROBLEM

More information

All in One First Strand cdna Synthesis Kit For reliable first strand cdna synthesis from all RNA sources

All in One First Strand cdna Synthesis Kit For reliable first strand cdna synthesis from all RNA sources All in One First Strand cdna Synthesis Kit For reliable first strand cdna synthesis from all RNA sources Cat. No. AORT 0020 Cat. No. AORT 0050 (20 synthesis reactions) (50 synthesis reactions) User Manual

More information

Essentials of Real Time PCR. About Sequence Detection Chemistries

Essentials of Real Time PCR. About Sequence Detection Chemistries Essentials of Real Time PCR About Real-Time PCR Assays Real-time Polymerase Chain Reaction (PCR) is the ability to monitor the progress of the PCR as it occurs (i.e., in real time). Data is therefore collected

More information

CompleteⅡ 1st strand cdna Synthesis Kit

CompleteⅡ 1st strand cdna Synthesis Kit Instruction Manual CompleteⅡ 1st strand cdna Synthesis Kit Catalog # GM30401, GM30402 Green Mountain Biosystems. LLC Web: www.greenmountainbio.com Tel: 800-942-1160 Sales: Sales@ greenmountainbio.com Support:

More information

PrimePCR Assay Validation Report

PrimePCR Assay Validation Report Gene Information Gene Name Gene Symbol Organism Gene Summary Gene Aliases RefSeq Accession No. UniGene ID Ensembl Gene ID papillary renal cell carcinoma (translocation-associated) PRCC Human This gene

More information

Table of Contents. I. Description... 2. II. Kit Components... 2. III. Storage... 2. IV. 1st Strand cdna Synthesis Reaction... 3

Table of Contents. I. Description... 2. II. Kit Components... 2. III. Storage... 2. IV. 1st Strand cdna Synthesis Reaction... 3 Table of Contents I. Description... 2 II. Kit Components... 2 III. Storage... 2 IV. 1st Strand cdna Synthesis Reaction... 3 V. RT-PCR, Real-time RT-PCR... 4 VI. Application... 5 VII. Preparation of RNA

More information

qstar mirna qpcr Detection System

qstar mirna qpcr Detection System qstar mirna qpcr Detection System Table of Contents Table of Contents...1 Package Contents and Storage Conditions...2 For mirna cdna synthesis kit...2 For qstar mirna primer pairs...2 For qstar mirna qpcr

More information

All-in-One mirna qrt-pcr Reagent Kits For quantitative detection of mature mirna

All-in-One mirna qrt-pcr Reagent Kits For quantitative detection of mature mirna All-in-One mirna qrt-pcr Reagent Kits For quantitative detection of mature mirna All-in-One TM mirna First-Strand cdna Synthesis Kit AMRT-0020 (20 RT reactions), AMRT-0060 (60 RT reactions) Used in combination

More information

RT31-020 20 rxns. RT31-100 100 rxns TRANSCRIPTME Enzyme Mix (1) 40 µl 2 x 50 µl 5 x 40 µl

RT31-020 20 rxns. RT31-100 100 rxns TRANSCRIPTME Enzyme Mix (1) 40 µl 2 x 50 µl 5 x 40 µl Components RT31-020 20 rxns RT31-050 50 rxns RT31-100 100 rxns TRANSCRIPTME Enzyme Mix (1) 40 µl 2 x 50 µl 5 x 40 µl 2x RT Master Mix (2) 200 µl 2 x 250 µl 5 x 200 µl RNase H (E. coli) 20 µl 2 x 25 µl

More information

Reconstituting and Diluting Primers and TaqMan Probes

Reconstituting and Diluting Primers and TaqMan Probes Reconstituting and Diluting Primers and TaqMan Probes Introduction TaqMan primers are commonly shipped in a lyophilized state. TaqMan probes are sometimes shipped in the lyophilized state, but more often

More information

HiPer RT-PCR Teaching Kit

HiPer RT-PCR Teaching Kit HiPer RT-PCR Teaching Kit Product Code: HTBM024 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 4 hours Agarose Gel Electrophoresis: 45 minutes Storage Instructions: The

More information

Oligo d(t) Primers. Product Specification. Reverse Transcriptase Primers, cdna Cloning Primers, T7 RNA Amplification Primers

Oligo d(t) Primers. Product Specification. Reverse Transcriptase Primers, cdna Cloning Primers, T7 RNA Amplification Primers Product Specification Reverse Transcriptase Primers, cdna Cloning Primers, T7 RNA Amplification Primers Oligo d(t) Primers Shipped at ambient temperature. Store at -20 o C For research use only. Not for

More information

mircute mirna qpcr Detection Kit (SYBR Green)

mircute mirna qpcr Detection Kit (SYBR Green) mircute mirna qpcr Detection Kit (SYBR Green) For detection of mirna using real-time RT-PCR (SYBR Green I) www.tiangen.com QP110302 mircute mirna qpcr Detection Kit (SYBR Green) Kit Contents Cat. no. FP401

More information

quantitative real-time PCR, grain, simplex DNA extraction: PGS0426 RT-PCR: PGS0494 & PGS0476

quantitative real-time PCR, grain, simplex DNA extraction: PGS0426 RT-PCR: PGS0494 & PGS0476 BioScience quantitative real-time PCR, grain, simplex DNA extraction: PGS0426 RT-PCR: PGS0494 & PGS0476 This method describes a Real-time semi-quantitative TaqMan PCR procedure for the determination of

More information

Mir-X mirna First-Strand Synthesis and SYBR qrt-pcr

Mir-X mirna First-Strand Synthesis and SYBR qrt-pcr User Manual Mir-X mirna First-Strand Synthesis and SYBR qrt-pcr User Manual United States/Canada 800.662.2566 Asia Pacific +1.650.919.7300 Europe +33.(0)1.3904.6880 Japan +81.(0)77.543.6116 Clontech Laboratories,

More information

User Manual/Hand book. qpcr mirna Arrays ABM catalog # MA003 (human) and MA004 (mouse)

User Manual/Hand book. qpcr mirna Arrays ABM catalog # MA003 (human) and MA004 (mouse) User Manual/Hand book qpcr mirna Arrays ABM catalog # MA003 (human) and MA004 (mouse) Kit Components Cat. No. MA003...Human Whole Genome mirna qpcr Profiling Kit (-20 C) The following components are sufficient

More information

Catherine Technical Product Specialist

Catherine Technical Product Specialist 量 PCR 理 Catherine Technical Product Specialist Traditional Quantitative System StepOne Real-Time PCR System Northern Blots Analysis Competitive PCR RT-PCR followed by South blots Disadvantages Labor intensive,

More information

Beginner s Guide to Real-Time PCR

Beginner s Guide to Real-Time PCR Beginner s Guide to Real-Time PCR 02 Real-time PCR basic principles PCR or the Polymerase Chain Reaction has become the cornerstone of modern molecular biology the world over. Real-time PCR is an advanced

More information

Real-Time PCR Set-up Instructions (for abm Reverse Transcription Products)

Real-Time PCR Set-up Instructions (for abm Reverse Transcription Products) Real-Time PCR Set-up Instructions (for abm Reverse Transcription Products) The following protocol is for real-time PCR using abm s EvaGreen qpcr MasterMix (Cat. No. MasterMix-R, -LR, -ic and -S). Please

More information

Epidemic Parotitis Virus Real Time RT-PCR Kit

Epidemic Parotitis Virus Real Time RT-PCR Kit Revision No.: ZJ0001 Issue Date: Aug 28th, 2007 Epidemic Parotitis Virus Real Time RT-PCR Kit Cat. No.: RR-0056-01 For Use with LightCycler2.0/LightCycler480 (Roche) Real Time PCR Systems For in vitro

More information

Novel In-process Monitoring Tools for Improving Accuracy and Reproducibility of RT-qPCR Results. Dirk Schacht, Global Product Manager, June 2015

Novel In-process Monitoring Tools for Improving Accuracy and Reproducibility of RT-qPCR Results. Dirk Schacht, Global Product Manager, June 2015 Novel In-process Monitoring Tools for Improving Accuracy and Reproducibility of RT-qPCR Results Dirk Schacht, Global Product Manager, June 2015 In-process monitoring tools for RT-qPCR 1 4-Part webinar

More information

All-in-One mirna qrt-pcr Detection System Handbook

All-in-One mirna qrt-pcr Detection System Handbook All-in-One mirna qrt-pcr Detection System Handbook For quantitative detection of mature mirna All-in-One mirna First-Strand cdna Synthesis Kit Cat. No. AMRT-0020 (20 mirna reverse transcription reactions)

More information

Absolute Quantification Getting Started Guide

Absolute Quantification Getting Started Guide 5 cdna Reverse Primer Oligo d(t) or random hexamer Synthesis of 1st cdna strand 3 5 cdna Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Absolute Quantification Getting Started Guide Introduction

More information

Protocol. Introduction to TaqMan and SYBR Green Chemistries for Real-Time PCR

Protocol. Introduction to TaqMan and SYBR Green Chemistries for Real-Time PCR Protocol Introduction to TaqMan and SYBR Green Chemistries for Real-Time PCR Copyright 2008, 2010 Applied Biosystems. All rights reserved. Ambion and Applied Biosystems products are for Research Use Only.

More information

FastLine cell cdna Kit

FastLine cell cdna Kit 1. FastLine cell cdna Kit For high-speed preparation of first-strand cdna directly from cultured cells without RNA purification www.tiangen.com RT100701 FastLine cell cdna Kit Cat. no. KR105 Kit Contents

More information

Speed Matters - Fast ways from template to result

Speed Matters - Fast ways from template to result qpcr Symposium 2007 - Weihenstephan Speed Matters - Fast ways from template to result March 28, 2007 Dr. Thorsten Traeger Senior Scientist, Research and Development - 1 - Overview Ạgenda Fast PCR The Challenges

More information

Put the odds in your favor

Put the odds in your favor Reverse Transcription Put the odds in your favor SuperScript III Reverse Transcriptase Reverse Transcription Engineered for performance SuperScript III Reverse Transcriptase (RT) is engineered for reduced

More information

AffinityScript QPCR cdna Synthesis Kit

AffinityScript QPCR cdna Synthesis Kit AffinityScript QPCR cdna Synthesis Kit INSTRUCTION MANUAL Catalog #600559 Revision C.01 For In Vitro Use Only 600559-12 LIMITED PRODUCT WARRANTY This warranty limits our liability to replacement of this

More information

RevertAid Premium First Strand cdna Synthesis Kit

RevertAid Premium First Strand cdna Synthesis Kit RevertAid Premium First Strand cdna Synthesis Kit #K1651, #K1652 CERTIFICATE OF ANALYSIS #K1651 Lot QUALITY CONTROL RT-PCR using 100 fg of control GAPDH RNA and GAPDH control primers generated a prominent

More information

MystiCq microrna cdna Synthesis Mix Catalog Number MIRRT Storage Temperature 20 C

MystiCq microrna cdna Synthesis Mix Catalog Number MIRRT Storage Temperature 20 C microrna cdna Synthesis Mix Catalog Number MIRRT Storage Temperature 20 C Product Description The microrna cdna Synthesis Mix has been designed to easily convert micrornas into cdna templates for qpcr

More information

Real-Time PCR Vs. Traditional PCR

Real-Time PCR Vs. Traditional PCR Real-Time PCR Vs. Traditional PCR Description This tutorial will discuss the evolution of traditional PCR methods towards the use of Real-Time chemistry and instrumentation for accurate quantitation. Objectives

More information

SYBR Green PCR Master Mix and SYBR Green RT-PCR Reagents Kit

SYBR Green PCR Master Mix and SYBR Green RT-PCR Reagents Kit USER GUIDE SYBR Green PCR Master Mix and SYBR Green RT-PCR Reagents Kit Catalog Number 4309155 (Master Mix) and 4306736 (RT-PCR Reagents Kit) Publication Part Number 4310251 Rev. G Revision Date September

More information

PrimePCR Assay Validation Report

PrimePCR Assay Validation Report Gene Information Gene Name sorbin and SH3 domain containing 2 Gene Symbol Organism Gene Summary Gene Aliases RefSeq Accession No. UniGene ID Ensembl Gene ID SORBS2 Human Arg and c-abl represent the mammalian

More information

Power SYBR Green PCR Master Mix and Power SYBR Green RT-PCR Reagents Kit

Power SYBR Green PCR Master Mix and Power SYBR Green RT-PCR Reagents Kit USER GUIDE Power SYBR Green PCR Master Mix and Power SYBR Green RT-PCR Reagents Kit Catalog Number 4368577, 4367659, 4367660, 4368706, 4368702, 4368708 (Master Mix) and 4368711 (RT-PCR Reagents Kit) Publication

More information

QPCR Applications using Stratagene s Mx Real-Time PCR Platform

QPCR Applications using Stratagene s Mx Real-Time PCR Platform QPCR Applications using Stratagene s Mx Real-Time PCR Platform Dan Schoeffner, Ph.D Field Applications Scientist Dan.Schoeffner@Stratagene.com Tech. Services 800-894-1304 Polymerase Chain Reaction Melt

More information

REAL TIME PCR SYBR GREEN

REAL TIME PCR SYBR GREEN REAL TIME PCR SYBR GREEN 1 THE PROBLEM NEED TO QUANTITATE DIFFERENCES IN mrna EXPRESSION SMALL AMOUNTS OF mrna LASER CAPTURE SMALL AMOUNTS OF TISSUE PRIMARY CELLS PRECIOUS REAGENTS 2 THE PROBLEM QUANTITATION

More information

TaqMan Fast Advanced Master Mix. Protocol

TaqMan Fast Advanced Master Mix. Protocol TaqMan Fast Advanced Master Mix Protocol For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. Information in this document is subject to change without notice. APPLIED

More information

Quantitative Telomerase Detection Kit (QTD Kit)

Quantitative Telomerase Detection Kit (QTD Kit) Quantitative Telomerase Detection Kit (QTD Kit) Catalog No. MT3010, MT3011, MT3012 For Research Use Only. Not for use in diagnostic procedures 1 Table of Contents 1. Introduction Background Product Overview

More information

Real time and Quantitative (RTAQ) PCR. so I have an outlier and I want to see if it really is changed

Real time and Quantitative (RTAQ) PCR. so I have an outlier and I want to see if it really is changed Real time and Quantitative (RTAQ) PCR or.. for this audience so I have an outlier and I want to see if it really is changed Nigel Walker, Ph.D. Laboratory of Computational Biology and Risk Analysis, Environmental

More information

MMLV High Performance Reverse Transcriptase

MMLV High Performance Reverse Transcriptase MMLV High Performance Reverse Transcriptase Cat. Nos. RT80110K and RT80125K Connect with Epicentre on our blog (epicentral.blogspot.com), Facebook (facebook.com/epicentrebio), and Twitter (@EpicentreBio).

More information

Absolute Quantitation Using Standard Curve Getting Started Guide

Absolute Quantitation Using Standard Curve Getting Started Guide Applied Biosystems 7900HT Fast Real-Time PCR System Absolute Quantitation Using Standard Curve Getting Started Guide Introduction Designing an AQ Experiment Preparing the Samples and Reaction Plate Creating

More information

Path-ID Multiplex One-Step RT-PCR Kit

Path-ID Multiplex One-Step RT-PCR Kit USER GUIDE Path-ID Multiplex One-Step RT-PCR Kit TaqMan probe-based multiplex one-step real-time RT-PCR detection of RNA targets Catalog Numbers 4428206, 4428207, 4440022 Publication Part Number 4440907

More information

Legionella pneumophila Detection Kit (Real-Time)

Legionella pneumophila Detection Kit (Real-Time) Unzipping Genes P r o d u c t I n f o r m a t i o n MBPCR028 Legionella pneumophila Detection Kit (Real-Time) Description: Legionella pneumophila is a gram-negative, non-encapsulated, aerobic bacillus

More information

TITRATION OF raav (VG) USING QUANTITATIVE REAL TIME PCR

TITRATION OF raav (VG) USING QUANTITATIVE REAL TIME PCR Page 1 of 5 Materials DNase digestion buffer [13 mm Tris-Cl, ph7,5 / 5 mm MgCl2 / 0,12 mm CaCl2] RSS plasmid ptr-uf11 SV40pA Forward primer (10µM) AGC AAT AGC ATC ACA AAT TTC ACA A SV40pA Reverse Primer

More information

Oligonucleotide Stability Study

Oligonucleotide Stability Study Oligonucleotide Stability Study IDT is performing a long term study of oligo stability. The study has been ongoing for 2 years and has tested oligo stability under various conditions. An overview of the

More information

Sample Preparation and Reaction Set-up

Sample Preparation and Reaction Set-up Sample Preparation and Reaction Set-up Important aspects of sample preparation and reaction setup Sampling Sample storage RNA / DNA Extraction and quality RNA / cdna / DNA storage Primer and probes formulation

More information

User Manual. Reference Gene Panel Mouse Probe protocol. Version 1.1 August 2014 For use in quantitative real-time PCR

User Manual. Reference Gene Panel Mouse Probe protocol. Version 1.1 August 2014 For use in quantitative real-time PCR User Manual Reference Gene Panel Mouse Probe protocol Version 1.1 August 2014 For use in quantitative real-time PCR Reference Gene Panel Mouse Table of contents Background 4 Assays included in the panel

More information

Blend Taq / Blend Taq -Plus-

Blend Taq / Blend Taq -Plus- Instruction manual Blend Taq and Blend Taq -Plus- 0810 F0938K Blend Taq / Blend Taq -Plus- BTQ-101 250 U 200 reactions BTQ-201 250 U 200 reactions Contents [1] Introduction

More information

RT-PCR: Two-Step Protocol

RT-PCR: Two-Step Protocol RT-PCR: Two-Step Protocol We will provide both one-step and two-step protocols for RT-PCR. We recommend the twostep protocol for this class. In the one-step protocol, the components of RT and PCR are mixed

More information

Real-time PCR of Type A A(H1N1)2009 viruses

Real-time PCR of Type A A(H1N1)2009 viruses Centre National de Référence Du Virus Influenzae Région Nord Sylvie van der Werf Dominique Rousset Vincent Enouf Real-time PCR of Type A A(H1N1)2009 viruses Ref: SOP/FluAswl/031209 2 INTRODUCTION This

More information

The AB7900Fast and Principles of Real Time PCR

The AB7900Fast and Principles of Real Time PCR The AB7900Fast and Principles of Real Time PCR Michael Tavaria, PhD Scientific Applications Specialist, Applied Biosystems, Australia and New Zealand Outline Chemistries 7900Fast Real Time PCR System Applications

More information

Relative Quantification Applied Biosystems 7300/7500 Real Time PCR System

Relative Quantification Applied Biosystems 7300/7500 Real Time PCR System 5 3 5 cdna Reverse Primer Oligo d(t) or random hexamer 3 5 cdna Getting Started Guide Relative Quantification Applied Biosystems 7300/7500 Real Time PCR System Introduction and Example RQ Experiment Designing

More information

Single Cell-to-CT Kit. Protocol

Single Cell-to-CT Kit. Protocol Single Cell-to-CT Kit Protocol For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. Information in this document is subject to change without notice. APPLIED BIOSYSTEMS

More information

Assay Assembly for Miniaturized Quantitative PCR in a 384-well Format using the Echo Liquid Handler Celeste Glazer, Sammy Datwani Labcyte Inc.

Assay Assembly for Miniaturized Quantitative PCR in a 384-well Format using the Echo Liquid Handler Celeste Glazer, Sammy Datwani Labcyte Inc. TM Application Note G102 Echo Liquid Handler Assay Assembly for Miniaturized Quantitative PCR in a 384-well Format using the Echo Liquid Handler Celeste Glazer, Sammy Datwani Labcyte Inc. Abstract Quantitative

More information

Brilliant III Ultra-Fast SYBR Green QRT-PCR Master Mix

Brilliant III Ultra-Fast SYBR Green QRT-PCR Master Mix Brilliant III Ultra-Fast SYBR Green QRT-PCR Master Mix Instruction Manual Catalog #600886 (single kit) #600887 (10-pack kit) Revision C Research Use Only. Not for Use in Diagnostic Procedures. 600886-12

More information

RNA Extraction and Quantification, Reverse Transcription, and Real-time PCR (q-pcr)

RNA Extraction and Quantification, Reverse Transcription, and Real-time PCR (q-pcr) RNA Extraction and Quantification, Reverse Transcription, and Real-time Preparation of Samples Cells: o Remove media and wash cells 2X with cold PBS. (2 ml for 6 well plate or 3 ml for 6cm plate) Keep

More information

ONLINE SUPPLEMENTAL MATERIAL. Allele-Specific Expression of Angiotensinogen in Human Subcutaneous Adipose Tissue

ONLINE SUPPLEMENTAL MATERIAL. Allele-Specific Expression of Angiotensinogen in Human Subcutaneous Adipose Tissue ONLINE SUPPLEMENTAL MATERIAL Allele-Specific Expression of Angiotensinogen in Human Subcutaneous Adipose Tissue Sungmi Park 1, Ko-Ting Lu 1, Xuebo Liu 1, Tapan K. Chatterjee 2, Steven M. Rudich 3, Neal

More information

Brilliant II QRT-PCR Master Mix with ROX, 1-Step

Brilliant II QRT-PCR Master Mix with ROX, 1-Step Brilliant II QRT-PCR Master Mix with ROX, 1-Step Instruction Manual Catalog #600838 (Brilliant II QRT-PCR High ROX Master Mix, 1-Step) #600837 (Brilliant II QRT-PCR Low ROX Master Mix, 1-Step) #600842

More information

Optimizing Real Time PCR: A Focused Approach for Exceptional Real-Time qpcr Results

Optimizing Real Time PCR: A Focused Approach for Exceptional Real-Time qpcr Results Optimizing Real Time PCR: A Focused Approach for Exceptional Real-Time qpcr Results Michael Wakem M.Sc Bio-Rad Laboratories Canada Ltd (800) 268-0213 Ext 3360 michael_wakem@bio-rad.com Overview Fundamentals

More information