Trichomonas Foetus DNA Test Kit
|
|
- Antony Manning
- 7 years ago
- Views:
Transcription
1 Trichomonas Foetus DNA Test Kit VetMAX -Gold Trich Detection Kit Catalog Number Pub. No Rev. A WARNING! Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves. Safety Data Sheets (SDSs) are available from Product information Name, intended use, and principle of the procedure The Trichomonas Foetus DNA Test Kit (VetMAX -Gold Trich Detection Kit) is a highly sensitive, qualitative, real-time PCR assay for detection of Tritrichomonas foetus DNA isolated from enriched smegma samples. Bovine trichomonosis is a sexually transmitted infection caused by T. foetus that results in significant monetary losses to the cattle industry in the United States and other parts of the world where open range management and natural breeding are practiced. T. foetus is a flagellated protozoan found in bovines that colonizes the vaginal, uterine, oviduct, and preputial epithelium, resulting in embryonic death, abortion, and infertility in the female. Although bulls are the main carriers of T. foetus, they remain asymptomatic for their entire lives. The VetMAX -Gold Trich Detection Kit enables detection of venereal trichomonosis in carrier bulls. The VetMAX -Gold Trich Detection Kit can be used to test pools of up to 5 samples for the presence of T. foetus. Pool size should be determined by the testing laboratory, based on the prevalence of bovine trichomonosis in the area from which the samples were collected. Pooled samples yielding a positive result should then be tested individually to determine the infection status of each animal in the positive pool. The assay is a single-well/tube, real-time PCR in which T. foetus DNA and Xeno Control DNA targets are amplified and detected in real time using fluorescent TaqMan probes (hydrolysis probe chemistry). The kit includes: T. foetus-xeno Control DNA Mix: serves as a positive control for the real-time PCR components, and it is also used to set the cycle threshold (C T ) for evaluating test results. Xeno Control DNA: serves as an internal control for the DNA isolation process, and it is also used to monitor for the presence of PCR inhibitors. T. foetus Primer Probe Mix for optimized multiplex PCR amplification of both Xeno Control DNA and T. foetus targets. Limitations Handle samples as recommended in Table 1 to prevent degradation of any T. foetus DNA that is present. Prepare pooled samples from no more than 5 individual samples. Pooling samples may cause loss of sensitivity of detection of infected animals with an individual C T of ³35. DNA extraction methods should yield DNA free of PCR inhibitors, which can prevent amplification of target DNA. Follow Good laboratory practices for PCR and RT-PCR on page 5 to prevent false positive amplifications due to contamination of test samples with PCR products. Kit contents and storage conditions Reagents for 100 real-time PCR tests (25 µl) are supplied. Component Volume Storage 2X qpcr Master Mix 1375 μl 30 C to 10 C T. foetus Primer Probe Mix 110 μl 30 C to 10 C Xeno Control DNA (10,000 copies/µl) 250 μl 30 C to 10 C T. foetus-xeno Control DNA Mix (1000 copies/µl) 80 μl 30 C to 10 C Nuclease-free Water 1.75 ml 30 C to +25 C Required materials not supplied Item Plates or tubes appropriate for the Applied Biosystems 7500 Fast Real-Time PCR System (96-well) Nuclease-free pipettes and filtered pipette tips Nuclease-free reagent tubes for preparing master mixes Real-time PCR thermal cycler 1X Phosphate Buffered Saline (PBS), ph 7.4 Source MicroAmp Optical 8-Cap Strip (Cat. no ), or equivalent MicroAmp Fast Optical 96-Well Reaction Plate with Barcode, 0.1-mL (Cat. nos , ), or equivalent. MicroAmp Optical Adhesive Film (Cat. nos , ), or equivalent MicroAmp Fast 8-Tube Strip, 0.1-mL (Cat. no ), or equivalent Precision Plate Holder for 0.1-mL tube strips (Cat. no ), or equivalent Major laboratory supplier (MLS) MLS Applied Biosystems 7500 Fast Real-Time PCR System (96-well), running SDS software v1.4 MLS InPouch TF Bovine BioMed Diagnostics (Cat. nos , ) 2 ice buckets: One for the PCR setup area where the master mix is prepared One for the area where DNA may be present MLS For Veterinary Use Only.
2 Isolate DNA from samples Table 1 Sample handling recommendations Step or process Transport/store samples Prepare cultured smegma samples (Optional) Prepare pooled cultured smegma samples Prepare mock-purified samples (for use in extraction control PCRs) Proposed DNA isolation method Required modifications to the DNA isolation method Recommendation 1. Transport inoculated InPouch media to the diagnostic lab, according to the manufacturer's instructions. 2. Incubate samples at 37 C for 24 hours. 1. Thoroughly mix the InPouch contents. 2. Transfer 1 ml from the pouch to a 1.5-mL tube. 3. Vigorously vortex the tube. 4. Use 300 μl of the cultured sample for DNA isolation. Pool up to 5 individual cultured smegma samples as described below [1]. 1. Thoroughly mix the individual cultured smegma sample, and create a 300-µL pool of cultured samples by combining equal volumes of individual samples. For example, transfer 60 μl of 5 individual samples to create one pool. 2. Briefly vortex the pooled cultures (properly sealed to prevent cross-contamination) to ensure sufficient mixing, and centrifuge briefly to collect the contents in the lower portion of the tube/plate. 3. Use 300 μl of the pooled smegma samples for each DNA purification, and process in the same way as individual samples. Prepare duplicate mock-purified samples, using 1X PBS as the starting material. Process mock-purified samples with the same DNA isolation method that is used for test samples. 5X MagMAX Pathogen RNA/DNA Extraction Kit (Cat. no ) or an equivalent DNA isolation method. Add 2 µl of undiluted Xeno Control DNA (20,000 copies) per purification to the lysis solution used for DNA purification. Add carrier RNA to the lysis solution according to the manufacturer's recommendation. Add carrier RNA in addition to Xeno Control DNA. Carrier RNA is provided in the 5X MagMAX Pathogen RNA/DNA Extraction Kit. [1] Pool size is determined by the testing laboratory. Perform real-time PCR 1 Determine the quantity of reactions and thaw the reagents a. Plan to include the following control reactions on each plate: Duplicate positive control reactions; use 8 µl of T. foetus-xeno Control DNA Mix (1000 copies/µl) per reaction. Duplicate no-template control (NTC) reactions; use Nuclease-free Water in place of sample DNA. b. Plan the plate layout so that the wells containing NTCs are located as far away as possible from positive controls and test samples to prevent accidental cross-contamination. c. Thaw PCR master mix reagents in one ice bucket and controls and samples in a separate ice bucket, gently vortex each tube to mix the contents thoroughly, then briefly centrifuge to collect the solution at the bottom of the tube. Keep the reagents on ice. 2 Prepare the real-time PCR master mix on ice Combine the following components for the number of reactions required plus 10% overage. Component Volume per reaction 2X qpcr Master Mix 12.5 μl T. foetus Primer Probe Mix 1.0 μl Nuclease-free Water 3.5 μl Total volume of real-time PCR master mix 17.0 μl 3 Set up the PCR reactions a. Dispense 17 µl of real-time PCR master mix to the appropriate wells of a PCR plate or PCR tubes on ice. b. Add the appropriate component for each reaction type, according to the following table: Reaction type Component Volume per reaction Test sample Sample DNA 8.0 μl NTC Nuclease-free Water 8.0 μl Positive control T. foetus-xeno Control DNA Mix (1000 copies/µl) 8.0 μl Extraction control Mock-purified PBS sample 8.0 μl 2 Trichomonas Foetus DNA Test Kit Instructions
3 3 Set up the PCR reactions (continued) c. Seal each plate or tube, mix, then centrifuge briefly to bring the contents to the bottom. 4 Set up and run the realtime PCR instrument a. Following the manufacturer s instructions, set up the run using the following parameters: Run mode: Standard 7500 Reaction volume: 25 µl ROX passive reference dye: Included in the 2X qpcr Master Mix TaqMan probe reporter dyes and quenchers: Target Reporter Quencher T. foetus DNA FAM dye [1] Eclipse Q Xeno Control DNA VIC dye [2] Eclipse Q [1] Absorbance maximum of 495 nm; emission maximum of 520 nm. [2] Absorbance maximum of 540 nm; emission maximum of 552 nm. b. Run the thermal cycler program and collect real-time amplification data during stage 2. Use the following thermal cycler settings: Stage Reps. Temp. Time Initial denaturation C 10 minutes Amplification C 55 C 15 seconds 45 seconds Data analysis Refer to your real-time PCR instrument user guide for instructions on data analysis, using the following method. Table 2 Data analysis Method Use the Control-Based Threshold setting for data analysis. Check the raw fluorescence data. Details 1. Select Manual C T. 2. Export DRn values for the positive control samples (T. foetus-xeno Control DNA Mix). 3. Average the FAM and VIC values (separately) for the DRn at cycle 40 for all replicates of the positive control reaction. 4. Set the threshold for the T. foetus (Trich) DNA target reactions at 10% of the average maximum fluorescence value of the T. foetus amplification signal in the positive control reactions (containing approximately 8000 copies of T. foetus-xeno Control DNA Mix per reaction). Example: If the average maximum fluorescence value for the T. foetus target in the positive control reactions is 0.15, set the T. foetus threshold at Repeat step 4 for the Xeno Control DNA target using a 10% threshold. Example: If the average maximum fluorescence value for the Xeno Control DNA target in the positive control reactions is 2.0, set the Xeno Control DNA threshold at 0.2. Verify that increased fluorescence seen in the normalized data is also evident without mathematical data processing. Interpretation of test results Table 3 Criteria for a valid real-time PCR run Reaction type C T value for T. foetus DNA C T value for Xeno Control DNA Positive control NTC 40 (no signal detected) [1] 40 (no signal detected) [1] Extraction control 40 (no signal detected) [1] [1] The run is invalid if the C T values for either T. foetus or Xeno Control DNA targets in the NTC are <40; see Troubleshooting on page 5. Trichomonas Foetus DNA Test Kit Instructions 3
4 Table 4 Interpretation of test results from individual samples C T value for T. foetus DNA C T value for Xeno Control DNA Interpretation < [1] T. foetus positive sample 40 (no signal detected) Xeno Control DNA C T shift 2.5 [2] T. foetus negative sample ³38 40 [2] Suspect result [2] [1] High levels of T. foetus DNA in the sample can reduce the signal from the Xeno Control DNA, resulting in a higher C T value for Xeno Control DNA in T. foetus-positive samples. See Troubleshooting on page 5. [2] See Table 5. Table 5 Assessment of suspect results (for individual samples only) Result Suspect result 1: The sample T. foetus C T value is <40 and the C T value for one or more NTCs or extraction controls is <40. Suspect result 2: A T. foetus (TF) sample is considered suspect if the TF C T value is 38 to 40. Action Repeat the extraction or real-time PCR. See No template control reaction on page 5. Analyze suspect DNA samples for the presence/absence of PCR inhibitors by calculating the Xeno Control DNA C T Shift: Xeno Control DNA C T Shift = SS XEC, where: SS = C T of Xeno Control DNA in the suspect sample XEC = Average C T of Xeno Control DNA in the extraction controls Workflow A Xeno Control DNA C T shift < Repeat the DNA purification of the original diagnostic sample in triplicate aliquots. 2. Repeat the real-time PCR with 8 μl of the purified DNA (for each triplicate aliquot). 3. Determine the number of samples with a C T value <40: 0 of 3: T. foetus negative. 1 of 3: Presumptive positive. ³2 of 3: T. foetus positive. Workflow B Xeno Control DNA C T shift ³ Repeat the real-time PCR with 2 μl of the suspect DNA sample. (PCR inhibitors may be present in the DNA.) If the T. foetus C T value is: <38: The sample is T. foetus positive; no further testing is required [1]. ³38: Continue with steps 2 5 of this workflow. 2. Dilute the original individual diagnostic sample 1:4 with 1X PBS. 3. Repeat the DNA purification of the 1:4 diluted sample in triplicate aliquots. 4. Repeat the real-time PCR with 8 μl of the purified DNA (for each triplicate aliquot). 5. Determine the number of samples with a T. foetus C T value <40: 0 of 3: T. foetus negative. 1 of 3: Presumptive positive. ³2 of 3: T. foetus positive. [1] The Xeno Control DNA C T value on diluted samples is not considered utilizing this workflow. Table 6 Interpretation of test results from pooled samples C T value for T. foetus DNA C T value for Xeno Control DNA Interpretation Action < 40 (signal detected) <40 (signal detected) T. foetus-positive pool 40 (no signal detected) Xeno Control DNA C T shift 2.5 T. foetus-negative pool None required Xeno Control DNA C T shift ³2.5 Suspect result Retest individual samples from the positive pool to determine which individual sample or samples caused the positive result. Interpret the retest results according to Table 4. Repeat the DNA purification with the individual cultured smegma samples, then repeat the real-time PCR with each DNA sample. Interpret the retest results according to Table 4. 4 Trichomonas Foetus DNA Test Kit Instructions
5 Troubleshooting Observation Possible cause Recommended action Positive control reaction T. foetus DNA no signal Xeno Control DNA no signal No template control reaction Signal detected (C T value is < 40) Extraction control reaction Signal detected (C T value is < 40) Test samples Xeno Control DNA no or low signal T. foetus DNA high signal Test samples Xeno Control DNA no signal T. foetus DNA no signal or suspect-range signal The T. foetus-xeno Control DNA Mix was improperly handled, resulting in DNA degradation. The 2X qpcr Master Mix was stored or handled improperly and it lost activity. The thermal cycler was not properly set up. The real-time PCR master mix was prepared incorrectly. There was contamination during the PCR. There was contamination during the DNA isolation or PCR. The Xeno Control DNA primers and probe are at limiting concentrations in the real-time PCR. High levels of T. foetus DNA in a sample can reduce amplification of the Xeno Control DNA. Use appropriate precautions against DNase contamination when handling the control DNAs. For example, wear clean gloves and use nuclease-free barrier pipette tips. Repeat the real-time PCR with fresh reagents. Check the thermal cycler settings. See Set up and run the realtime PCR instrument on page 3. Repeat the test with correctly prepared real-time PCR master mix. Repeat the real-time PCR with fresh reagents and freshly decontaminated pipettors. Set up the real-time PCR in an area separate from areas used for DNA isolation and PCR product analysis. Repeat the DNA isolation or real-time PCR with fresh reagents and freshly decontaminated pipettors. Set up the real-time PCR in an area separate from areas used for DNA isolation and PCR product analysis. No or low signal from Xeno Control DNA is expected in a reaction that has a strong signal for T. foetus DNA. Poor DNA recovery. Check the C T values of Xeno Control DNA in the mockpurified samples. A C T value ³ 38 in individual or pooled samples indicates that Xeno Control DNA was omitted or that DNA recovery was poor. Repeat the DNA purification of the original diagnostic sample. The DNA samples contain PCR inhibitors. See Table 5. Explanation of symbols The symbols present on the product label are explained in the following table. MANUFACTURER CATALOG NUMBER BATCH CODE SERIAL NUMBER USE BY CONSULT INSTRUCTIONS FOR USE CAUTION, CONSULT ACCOMPANYING DOCUMENTS UPPER AND LOWER LIMITS OF TEMPERATURE Good laboratory practices for PCR and RT-PCR When preparing samples for PCR or RT-PCR amplification: Wear clean gloves and a clean lab coat (not previously worn while handling amplified products or used during sample preparation). Change gloves whenever you suspect that they are contaminated. Maintain separate areas and dedicated equipment and supplies for: Sample preparation and reaction setup. Amplification and analysis of products. Do not bring amplified products into the reaction setup area. Open and close all sample tubes carefully. Avoid splashing or spraying samples. Keep reactions and components capped as much as possible. Use a positive-displacement pipettor or aerosol-resistant barrier pipette tips. Clean lab benches and equipment periodically with 10% bleach solution or DNAZap Solutions (Cat. no. AM9890). Trichomonas Foetus DNA Test Kit Instructions 5
6 Safety Specific chemical handling CAS Sodium Azide Chemical information Sodium azide may react with lead and copper plumbing to form highly explosive metal azides. Limited product warranty Life Technologies Corporation and/or its affiliate(s) warrant their products as set forth in the Life Technologies' General Terms and Conditions of Sale found on Life Technologies' website at If you have any questions, please contact Life Technologies at For Veterinary Use Only. Manufacturer's address: Life Technologies Corporation 2130 Woodward Street Austin, TX US Veterinary License No The information in this guide is subject to change without notice. DISCLAIMER LIFE TECHNOLOGIES CORPORATION AND/OR ITS AFFILIATE(S) DISCLAIM ALL WARRANTIES WITH RESPECT TO THIS DOCUMENT, EXPRESSED OR IMPLIED, INCLUDING BUT NOT LIMITED TO THOSE OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE, OR NON-INFRINGEMENT. TO THE EXTENT ALLOWED BY LAW, IN NO EVENT SHALL LIFE TECHNOLOGIES AND/OR ITS AFFILIATE(S) BE LIABLE, WHETHER IN CONTRACT, TORT, WARRANTY, OR UNDER ANY STATUTE OR ON ANY OTHER BASIS FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING BUT NOT LIMITED TO THE USE THEREOF. LIMITED USE LABEL LICENSE No. 460: PCR Veterinary Diagnostics: Notice to Purchaser: For veterinary diagnostic services including reporting the results of such services for a fee and for research purposes only. Human diagnostic uses require a separate license from Roche Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. InPouch is a trademark of VWR International, Inc. TaqMan is a registered trademark of Roche Molecular Systems, Inc., used under permission and license. Eclipse is a registered trademark of Elitech Holding BV. For support visit lifetechnologies.com/support or techsupport@lifetech.com lifetechnologies.com 18 September 2013
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
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
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.
RealStar HBV PCR Kit 1.0 11/2012
RealStar HBV PCR Kit 1.0 11/2012 RealStar HBV PCR Kit 1.0 For research use only! (RUO) Product No.: 201003 96 rxns INS-201000-GB-02 Store at -25 C... -15 C November 2012 altona Diagnostics GmbH Mörkenstraße
RealLine HCV PCR Qualitative - Uni-Format
Instructions for use PCR KIT FOR EXTRACTION OF RNA AND REAL TIME PCR DETECTION KIT FOR HEPATITIS C VIRUS RNA Research Use Only Qualitative Uni Format VBD0798 48 tests valid from: December 2013 Rev11122013
HBV Quantitative Real Time PCR Kit
Revision No.: ZJ0002 Issue Date: Aug 7 th, 2008 HBV Quantitative Real Time PCR Kit Cat. No.: HD-0002-01 For Use with LightCycler 1.0/LightCycler2.0/LightCycler480 (Roche) Real Time PCR Systems (Pls ignore
Sequencing Library qpcr Quantification Guide
Sequencing Library qpcr Quantification Guide FOR RESEARCH USE ONLY Introduction 3 Quantification Workflow 4 Best Practices 5 Consumables and Equipment 6 Select Control Template 8 Dilute qpcr Control Template
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
Applied Biosystems KRAS Mutation Analysis Reagents. Protocol
Applied Biosystems KRAS Mutation Analysis Reagents Protocol For Research Use Use Only. Not intended for any animal or human therapeutic or diagnostic use. Information in this document is subject to change
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...
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...
One Shot TOP10 Competent Cells
USER GUIDE One Shot TOP10 Competent Cells Catalog Numbers C4040-10, C4040-03, C4040-06, C4040-50, and C4040-52 Document Part Number 280126 Publication Number MAN0000633 Revision A.0 For Research Use Only.
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
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
Fluorescein Isothiocyanate (FITC)- conjugated Antibodies
USER GUIDE Fluorescein Isothiocyanate (FITC)- conjugated Antibodies Catalog Numbers R933-25, R953-25, R963-25 Document Part Number 25-0376 Publication Number MAN0000194 Revision 2.0 For Research Use Only.
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
OpenArray Sample Tracker Software
QUICK REFERENCE OpenArray Sample Tracker Software For QuantStudio 12K Flex OpenArray Sample Block and the OpenArray Real-Time PCR System Publication Part Number 4460657 Rev. C Revision Date May 2012 Contents
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...
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
qpcr Quantification Protocol Guide
qpcr Quantification Protocol Guide FOR RESEARCH USE ONLY Topics 3 Introduction 5 User-Supplied Consumables and Equipment 7 Select Template 8 Dilute qpcr Template 9 Dilute Libraries 10 Prepare Reaction
FOR REFERENCE PURPOSES
BIOO LIFE SCIENCE PRODUCTS FOR REFERENCE PURPOSES This manual is for Reference Purposes Only. DO NOT use this protocol to run your assays. Periodically, optimizations and revisions are made to the kit
High-Capacity cdna Reverse Transcription Kits For 200 and 1000 Reactions
High-Capacity cdna Reverse Transcription Kits For 200 and 1000 Reactions Protocol Copyright 2006,. All rights reserved. For Research Use Only. Not for use in diagnostic procedures. Information in this
Applied Biosystems 3500/3500xL Genetic Analyzer
QUICK REFERENCE Applied Biosystems 3500/3500xL Genetic Analyzer with 3500 Series Data Collection Software 3 Catalog Number 4405186, 4405187 Pub. No. 100026299 Rev. A Note: For safety and biohazard guidelines,
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
Intended Use: The kit is designed to detect the 5 different mutations found in Asian population using seven different primers.
Unzipping Genes MBPCR014 Beta-Thalassemia Detection Kit P r o d u c t I n f o r m a t i o n Description: Thalassemia is a group of genetic disorders characterized by quantitative defects in globin chain
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
Quantifiler HP and Trio DNA Quantification Kits User Guide
USER GUIDE Quantifiler HP and Trio DNA Quantification Kits User Guide For use with: Quantifiler HP DNA Quantification Kit (Cat. no. 4482911) Quantifiler Trio DNA Quantification Kit (Cat. no. 4482910) Publication
Quantifiler Human DNA Quantification Kit Quantifiler Y Human Male DNA Quantification Kit
Product Bulletin Human Identification Quantifiler Human DNA Quantification Kit Quantifiler Y Human Male DNA Quantification Kit The Quantifiler kits produce reliable and reproducible results, helping to
UltraClean Soil DNA Isolation Kit
PAGE 1 UltraClean Soil DNA Isolation Kit Catalog # 12800-50 50 preps New improved PCR inhibitor removal solution (IRS) included Instruction Manual (New Alternative Protocol maximizes yields) Introduction
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
Factors Influencing Multiplex Real-Time PCR
APPLICATION NOTE Multiplex Real-Time PCR Factors Influencing Multiplex Real-Time PCR Introduction Multiplex PCR is the simultaneous amplification of more than one target sequence in a single reaction [1].
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
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
PCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System) 50 preps Product #60200
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com PCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System)
Megaplex Pools For microrna Expression Analysis
Megaplex Pools For microrna Expression Analysis Protocol Copyright 2008, 2010 Applied Biosystems. All rights reserved. For Research Use Only. Not for use in diagnostic procedures. Information in this document
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
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
TaqMan Copy Number Assays. Protocol
TaqMan Copy Number Assays 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. Life Technologies
HBV PCR detection Kit USER MANUAL
For professional use only HBV PCR detection Kit (PREP-NA DNA/RNA Extraction Kit included) USER MANUAL "DNA-Technology, Research & Production" LLC Russia, 142281, Moscow Region, Protvino, 2 Zheleznodorozhnaya
ID kit. imegen Anchovies II. and E. japonicus) DNA detection by. User manual. Anchovies species (E. encrasicolus. sequencing.
User manual imegen Anchovies II ID kit Anchovies species (E. encrasicolus and E. japonicus) DNA detection by sequencing Reference: Made in Spain The information in this guide is subject to change without
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
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
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
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
TaqMan Mutation Detection Assay
PROTOCOL TaqMan Mutation Detection Assay Competitive Allele-Specific TaqMan PCR Publication Part Number 4467011 Rev. B Revision Date April 2012 For Research Use Only. Not intended for any animal or human
DNA Analyst Training Laboratory Training Manual Protocol 2.02 Clean Technique
DNA Analyst Training Laboratory Training Manual Protocol 2.02 Clean Technique This laboratory protocol (or part thereof) has been provided as an example of a laboratory SOP, courtesy of the Illinois State
Real-time PCR: Understanding C t
APPLICATION NOTE Real-Time PCR Real-time PCR: Understanding C t Real-time PCR, also called quantitative PCR or qpcr, can provide a simple and elegant method for determining the amount of a target sequence
amplification tech Optical Design of CFX96 Real-Time PCR Detection System Eliminates the Requirement of a Passive Reference Dye
amplification tech note 6047 Optical Design of CFX96 Real-Time PCR Detection System Eliminates the Requirement of a Passive Reference Dye Liz Jordan and Richard Kurtz, Gene Expression Division, io-rad
PATHOGEN DETECTION SYSTEMS BY REAL TIME PCR. Results Interpretation Guide
PATHOGEN DETECTION SYSTEMS BY REAL TIME PCR Results Interpretation Guide Pathogen Detection Systems by Real Time PCR Microbial offers real time PCR based systems for the detection of pathogenic bacteria
ExpressArt Bacterial H-TR cdna synthesis kit. With extreme selectivity against rrnas
ExpressArt Bacterial H-TR cdna synthesis kit With extreme selectivity against rrnas suitable for 2 to 4 µg total RNA Catalogue No. 8004-A30 (30 rxns) Reagents Materials are provided for 30 cdna synthesis
PNA BRAF Mutation Detection Kit
- PNA BRAF Mutation Detection Kit Catalog Number KA2102 50 tests/kit Version: 01 Intended for research use only www.abnova.com Introduction and Background Intended use The PNA BRAF Mutation Detection Kit
Mouse IgM ELISA. Cat. No. KT-407 K-ASSAY. For the quantitative determination of IgM in mouse biological samples. For Research Use Only. 1 Rev.
K-ASSAY Mouse IgM ELISA For the quantitative determination of IgM in mouse biological samples Cat. No. KT-407 For Research Use Only. 1 Rev. 072309 K-ASSAY PRODUCT INFORMATION Mouse IgM ELISA Cat. No. KT-407
Plexor Systems Instrument Setup and Data Analysis for the Applied Biosystems 7300 and 7500 Real-Time PCR Systems
Technical Manual Plexor Systems Instrument Setup and Data Analysis for the Applied Biosystems 7300 and 7500 Real-Time PCR Systems INSTRUCTIONS FOR USE OF PRODUCTS A4011, A4021, A4031, A4041, A4051 AND
Platinum Multiplex PCR Master Mix
USER GUIDE Publication Part Number 4463722 Rev. A Revision Date April 2011 For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. Information in this document is subject
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.
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
altona altona RealStar Clostridium difficile PCR Kit 1.0 always a drop ahead. 11/2012 altona Diagnostics GmbH Moerkenstr. 12 22767 Hamburg Germany
altona DIAGNOSTICS altona DIAGNOSTICS RealStar Clostridium difficile PCR Kit 1.0 11/2012 altona Diagnostics GmbH Moerkenstr. 12 22767 Hamburg Germany phone +49 40 548 06 76-0 fax +49 40 548 06 76-10 e-mail
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:
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
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
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
Genomic DNA Extraction Kit INSTRUCTION MANUAL
Genomic DNA Extraction Kit INSTRUCTION MANUAL Table of Contents Introduction 3 Kit Components 3 Storage Conditions 4 Recommended Equipment and Reagents 4 Introduction to the Protocol 4 General Overview
Taqman TCID50 for AAV Vector Infectious Titer Determination
Page 1 of 8 Purpose: To determine the concentration of infectious particles in an AAV vector sample. This process involves serial dilution of the vector in a TCID50 format and endpoint determination through
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
TaqMan Salmonella Enteritidis Detection Kit. Egg and Environmental Testing Protocol
TaqMan Salmonella Enteritidis Detection Kit Egg and Environmental Testing Protocol Qualitative real-time PCR assay for the detection of Salmonella Enteritidis from pre-enriched poultry eggs and environmental
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
UltraClean Forensic DNA Isolation Kit (Single Prep Format)
UltraClean Forensic DNA Isolation Kit (Single Prep Format) Catalog No. Quantity 14000-10 10 preps 14000-S 1 prep Instruction Manual Please recycle Version: 10302012 1 Table of Contents Introduction...
Creatine Kinase (CK) Enzymatic Assay Kit Manual Catalog #: 3460-07
Creatine Kinase (CK) Enzymatic Assay Kit Manual Catalog #: 3460-07 TABLE OF CONTENTS GENERAL INFORMATION... 2 Product Description... 2 Procedure Overview... 2 Kit Contents, Storage and Shelf Life... 3
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
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,
Agencourt RNAdvance Blood Kit for Free Circulating DNA and mirna/rna Isolation from 200-300μL of Plasma and Serum
SUPPLEMENTAL PROTOCOL WHITE PAPER Agencourt RNAdvance Blood Kit for Free Circulating DNA and mirna/rna Isolation from 200-300μL of Plasma and Serum Bee Na Lee, Ph.D., Beckman Coulter Life Sciences Process
Application Guide... 2
Protocol for GenomePlex Whole Genome Amplification from Formalin-Fixed Parrafin-Embedded (FFPE) tissue Application Guide... 2 I. Description... 2 II. Product Components... 2 III. Materials to be Supplied
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
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
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
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
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
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
ncounter Gene Expression Assay Manual Total RNA and Cell Lysate Protocols
ncounter Gene Expression Assay Manual Total RNA and Cell Lysate Protocols v.20090807 For research use only. Not for use in diagnostic procedures. Limited License Subject to the terms and conditions of
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
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
Highly specific and sensitive quantitation
PRODUCT ULLETIN SYR Select Master Mix SYR Select Master Mix Highly specific and sensitive quantitation SYR Select Master Mix offers advanced performance at an affordable price. SYR Select Master Mix is
empcr Amplification Method Manual - Lib-A
GS Junior Titanium Series May 2010 (Rev. April 2011) For life science research only. Not for use in diagnostic procedures. 1. WORKFLOW The emulsion-based clonal amplification (empcr amplification) of a
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,
BacReady TM Multiplex PCR System
BacReady TM Multiplex PCR System Technical Manual No. 0191 Version 10112010 I Description.. 1 II Applications 2 III Key Features.. 2 IV Shipping and Storage. 2 V Simplified Procedures. 2 VI Detailed Experimental
Aurora Forensic Sample Clean-up Protocol
Aurora Forensic Sample Clean-up Protocol 106-0008-BA-D 2015 Boreal Genomics, Inc. All rights reserved. All trademarks are property of their owners. http://www.borealgenomics.com support@borealgenomics.com
UltraClean PCR Clean-Up Kit
UltraClean PCR Clean-Up Kit Catalog No. Quantity 12500-50 50 Preps 12500-100 100 Preps 12500-250 250 Preps Instruction Manual Please recycle Version: 02212013 1 Table of Contents Introduction... 3 Protocol
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
IgM ELISA. For the quantitative determination of IgM in human serum and plasma. For Research Use Only. Not For Use In Diagnostic Procedures.
IgM ELISA For the quantitative determination of IgM in human serum and plasma For Research Use Only. Not For Use In Diagnostic Procedures. Please read carefully due to Critical Changes, e.g., Calibrator
50 g 650 L. *Average yields will vary depending upon a number of factors including type of phage, growth conditions used and developmental stage.
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Phage DNA Isolation Kit Product # 46800, 46850 Product Insert
Data Sheet. PD-L1:B7-1[Biotinylated] Inhibitor Screening Assay Kit Catalog # 72026 Size: 96 reactions
Data Sheet PD-L1:B7-1[Biotinylated] Inhibitor Screening Assay Kit Catalog # 72026 Size: 96 reactions BACKGROUND: There have been a number of studies defining a role for PD-L1 in the regulation of immune
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
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
Absolute Quantitation Using Standard Curve 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 Quantitation Using Standard Curve Getting Started
RIBOPROTECT. RNase Inhibitor RT33-020, RT33-100
RIBOPROTECT RT33-020, RT33-100 RT33-020, RT33-100 RIBOPROTECT The RIBOPROTECT is a recombinant protein isolated and purified from Escherichia coli. It inhibits ribonuclease (RNase) activity of enzymes
PicoMaxx High Fidelity PCR System
PicoMaxx High Fidelity PCR System Instruction Manual Catalog #600420 (100 U), #600422 (500 U), and #600424 (1000 U) Revision C Research Use Only. Not for Use in Diagnostic Procedures. 600420-12 LIMITED
GeneXpert Technology. Indira Soundiram 2012
GeneXpert Technology Indira Soundiram 2012 A Better Way to Platform Design GeneXpert Infinity-48 GeneXpert Module GX-I GX-II GX-IV GX-XVI 2 Defining Molecular Diagnostics Any Test Any Time Any Sample Any
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...
TaqMan MicroRNA Cells-to-CT Kit (P/N 4391848, 4391996)
TaqMan MicroRNA Cells-to-CT Kit (P/N 4391848, 4391996) Protocol I. Introduction....................................................... 1 A. Product Description and Background B. Materials Provided with
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