Put the odds in your favor

Save this PDF as:
 WORD  PNG  TXT  JPG

Size: px
Start display at page:

Download "Put the odds in your favor"

Transcription

1 Reverse Transcription Put the odds in your favor SuperScript III Reverse Transcriptase

2 Reverse Transcription Engineered for performance SuperScript III Reverse Transcriptase (RT) is engineered for reduced RNase H activity and high thermostability (Figure 1), and delivers robust first-strand synthesis with both high cdna yield and ultimate sensitivity, whatever the size of your mrna. SuperScript III RT exhibits reduced RNase H activity, for greater production of full-length cdna and more complete gene representation (Figure 2). SuperScript III RT has also been engineered for high thermostability and has a half-life of 220 min at 50 C. This increases its ability to process RNA with secondary structure. More representative full-length cdna Increased cdna yields Performance-based evolution of SuperScript III Reverse Transcriptase SuperScript III Reverse Transcriptase is ideal for RT-PCR of a specific gene or for generating cdna from a total or poly(a) + RNA sample. It synthesizes a complementary DNA strand from single-stranded RNA or DNA, or an RNA:DNA hybrid. SuperScript Reverse Transcriptase SuperScript II Reverse Transcriptase SuperScript III SuperScript II MMLV M M SuperScript III Reverse Transcriptase Reduced RNase H activity Increased thermostability Figure 1. Performance-based evolution of SuperScript III Reverse Transcriptase. To meet the growing need for sensitivity, we developed SuperScript III Reverse Transcriptase, the latest-generation RT enzyme that provides improved performance in a variety of gene expression applications. Figure 2. SuperScript III RT generates the highest yields of full-length cdna. The autoradiograph shows 32 P-labeled cdna synthesized from 0.25 µg of mixed RNA, consisting of 1.35, 2.4, 4.4, 7.5, and 9.5 kb fragments, with 200 units of each RT at various temperatures ( C). Lane M is 32 P-labeled 1 Kb DNA Ladder. For up-to-date information on SuperScript III reverse transcription tools, visit

3 Overview: one-step and two-step RT-PCR There are two approaches for performing RT-PCR: one-step and Table 1. Comparison of one-step and two-step RT-PCR. two-step RT-PCR (Table 1). Two-step RT-PCR One-step RT-PCR One-step RT-PCR combines the first-strand cdna synthesis (reverse transcription) reaction and PCR reaction in the same tube, simplifying reaction setup and reducing the possibility of contamination. One-step RT-PCR allows easier processing of large numbers of First-strand cdna priming Choice of amplification reagents Oligo(dT) primers Random primers Sequence-specific primers Sequence-specific primers samples, and helps minimize carryover contamination, since tubes Fewer pipetting steps are not opened between cdna synthesis and amplification. By amplifying the entire cdna sample, one-step RT-PCR can provide greater sensitivity from as little as 0.01 pg total RNA. One-step Analysis of multiple cdnas from a sample Ability to optimize reactions allow for the use of sequence-specific primers only. Flexibility Two-step PCR begins with the reverse transcription of either Convenience total RNA or poly(a) + RNA into cdna using a reverse transcriptase. Following first-strand synthesis, the cdna is transferred to a separate tube for the PCR step. Two-step RT-PCR is useful for detecting multiple messages from a single RNA sample. You ll get greater flexibility when choosing primers and polymerase than with one-step RT-PCR systems. When performing two-step RT-PCR, you have the option SuperScript III Reverse Transcriptase 1 ng 10 ng 100 ng 1,000 ng Transcriptor reverse transcriptase 1 ng 10 ng 100 ng 1,000 ng of using either oligo(dt), random hexamer, or sequence-specific primers, and then PCR is performed with either Platinum Taq DNA Polymerase, Platinum Taq DNA Polymerase High Fidelity, or your choice of PCR enzyme. One-step RT-PCR The SuperScript III One-Step RT-PCR System with Platinum Taq DNA Polymerase delivers increased cdna yields, high sensitivity, and full-length transcripts in a convenient format (Figure 3). The high thermostability of SuperScript III RT combined with the Figure 3. Compared to Transcriptor reverse transcriptase (Roche), SuperScript III Reverse Transcriptase produces more full-length cdna and more representative cdna from endpoint RT-PCR. Yields of cdna are also higher with SuperScript III RT, increasing sensitivity and producing enough cdna for multiple downstream applications. Lane 1: GAPDH, 555 bp; lane 2: β-actin, 1,026 bp; lane 3: LAM, 1,490 bp; lane 4: BACH, 2,000 bp; lane 5: BF, 2.4 kb; lane 6: ACTR, 3,009 bp; lane 7: HTF4, 3.6 kb; lane 8: VIN, 4,497 bp; lane 9: MLL, 5,678 bp; lane 10: AKAP, 7,013 bp; lane 11: DNCH, 7,818 bp; lane 12: FIB, 9.4 kb; lane 13: MLL, 10,098 bp. specificity of Platinum Taq DNA Polymerase increases priming specificity, increases product yields, and identifies a larger range of targets.

4 Two-step RT-PCR The SuperScript III First-Strand Synthesis System for RT-PCR delivers increased cdna yields, high sensitivity, and full-length transcripts in a convenient format. You get all the components you need for successful first-strand cdna synthesis, saving you time and ensuring your success with every experiment. SuperScript III First-Strand Synthesis SuperMix The SuperMix format uses just two tubes, a 2x Reaction Mix and a SuperScript III Enzyme Mix, to provide simple, time-saving reaction setup. An optional annealing buffer is also provided. RNA priming strategies First-strand cdna synthesis reactions can use sequence-specific primers (SSPs), oligo(dt) primers, random primers, or a combination of primers. The RNA priming strategy can have a significant impact on cdna synthesis efficiency, consistency, and cdna yields. While each primer type has its benefits and drawbacks, individual RNA targets may respond differently to one RT primer choice over another (Table 2). Using a combination of random and oligo(dt) primers in reverse transcription can sometimes increase data quality by combining the benefits of both primer types. SuperScript III First-Strand Synthesis System for RT-PCR This system for RT-PCR provides all the necessary components for synthesis of high-quality first-strand cdna in separate tubes, to allow maximum optimization of cdna synthesis. Table 2. Priming strategies for cdna synthesis. cdna yield Specificity Multiple targets Difficult targets Bacterial RNA Sequence-specific primers NA Oligo(dT) primers NA Random primers

5 SuperScript VILO cdna Synthesis Kit The SuperScript VILO cdna Synthesis Kit is designed to increase cdna yields and improve the dynamic range of your qrt-pcr assays. This means that you obtain the same relative representation in your cdna regardless of the abundance level of your gene of interest. Reduce bias generated from low- or high-abundance genes Obtain reliable analysis of more genes Archive cdna for future studies Reduce bias in your reverse transcriptase (RT) reactions Obtain consistent and accurate results with unprecedented linearity across the broadest range of input material when you use the SuperScript VILO cdna Synthesis Kit (Figure 4). Because the bias toward abundant gene products is reduced, data derived from comparing the gene of interest and reference genes are more accurate. Reliable analysis of more genes The SuperScript VILO cdna Synthesis Kit, containing SuperScript III and a proprietary helper protein, minimizes carryover of PCR inhibitors, allowing you to add greater amounts of cdna to the qrt-pcr experiments without inhibiting downstream steps. This enables the analysis of both low- and high-abundance genes, regardless of their expression levels or the amount of RNA used in the reaction (Figure 5) C t ,000 10, ,000 1,000,000 10,000,000 Input RNA quantity (pg) Figure 4. The SuperScript VILO cdna Synthesis Kit provides reliable performance across an extended linear range of RNA input. Serial dilutions of total RNA from HeLa cells were reverse-transcribed using the SuperScript VILO cdna Synthesis Kit, followed by triplicate qpcr reactions using human β-actin TaqMan assays (Applied Biosystems) with the EXPRESS qpcr SuperMix Universal (Invitrogen) on an ABI PRISM 7900HT Sequence Detection System. The standard curve shows that even outside the recommended input of 1 pg to 2.5 µg, the data generated by the kit exhibit a coefficient of correlation of for up to 5 µg of input RNA. Rn Cycle Figure 5. The SuperScript VILO cdna Synthesis Kit is robust across an extended linear range of RNA input. The amplification plot illustrates that the triplicates are aligned across all dilutions, demonstrating the robustness of the SuperScript VILO cdna Synthesis Kit over a broad linear range of RNA input. This means that you can normalize your lower-abundance genes to your reference genes without worrying about potential variation of RT efficiency at different RNA input levels.

6 Archive cdna for future studies 30,000 25,000 25,687 The SuperScript VILO cdna Synthesis Kit, which includes SuperScript III RT, RNaseOUT RNase inhibitor, and a proprietary helper protein, generates far greater yields of cdna (Figure 6). The extra cdna product can be archived for subsequent applica- Radioactivity (cpm) 20,000 15,000 10,000 5,000 9,310 2,965 tions, and you also have the option of increasing the amount of cdna in your experiment for improved sensitivity (up to 4-fold) 0 SSIII qrt-pcr VILO RT system MMLV without the fear of inhibiting the reaction. Figure 6. The SuperScript VILO cdna Synthesis Kit exhibits less PCR inhibition for greater sensitivity in your real-time PCR application. Three different kits were compared for cdna yield, using 32 P-dCTP incorporation and the same amount of starting HeLa RNA: the Invitrogen SuperScript III First-Strand Synthesis SuperMix for qrt-pcr ( SSIII qrt-pcr ), the new SuperScript VILO cdna Synthesis Kit ( VILO ), and MMLV. The VILO kit shows 3- to 4-fold greater cdna yield than the other Invitrogen kit and 8- to 9-fold greater yield than MMLV. Ordering information Product Quantity Cat. No. SuperScript VILO cdna Synthesis Kit 50 x 20 µl rxns 250 x 20 µl rxns Learn more at SuperScript III One-Step RT-PCR System with Platinum Taq DNA Polymerase SuperScript III One-Step RT-PCR System with Platinum Taq High Fidelity 25 rxns 100 rxns 25 rxns 100 rxns SuperScript III First-Strand Synthesis SuperMix 50 rxns SuperScript III First-Strand Synthesis System for RT-PCR 50 rxns To learn more about one-step and two-step RT-PCR, visit SuperScript III Reverse Transcriptase Learn more at 2,000 units 10,000 units 4 x 10,000 units For research use only. Not intended for any animal or human therapeutic or diagnostic use, unless otherwise stated Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. These products may be covered by one or more Limited Use Label Licenses (see Invitrogen catalog or By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. B

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

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

RNA-related Products

RNA-related Products RNA-related Products TranscriptME RNA kit: Ideal choice for obtaining high yields of full-length cdna for RT-qPCR assays Suitable for as low RNA amount as 10 pg p p Convenient, reliable and cost-effective

More information

RNA-related Products

RNA-related Products RNA-related Products TRANSCRIPTME RNA kit: Ideal choice for obtaining high yields of full-length cdna for RT-qPCR assays Suitable for as low RNA amount as 10 pg p p Convenient, reliable and cost-effective

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

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

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

Reverse Transcription System

Reverse Transcription System TECHNICAL BULLETIN Reverse Transcription System Instruc ons for use of Product A3500 Revised 1/14 TB099 Reverse Transcription System All technical literature is available on the Internet at: www.promega.com/protocols/

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

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

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

GoTaq Amplification Maximum performance, flexibility, control and convenience

GoTaq Amplification Maximum performance, flexibility, control and convenience GoTaq Amplification Maximum performance, flexibility, control and convenience GoTaq products and formulations designed to improve your amplification needs Reverse Transcription GoScript Reverse Transcriptase

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

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

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

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

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

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

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

PCR and qpcr solutions

PCR and qpcr solutions A T C C C G C A T C C C G C A T C C C G C A T C C C G C A T C C G A T A T C C C G C A T C C C G C A T C C C G C A T C C C G C A T C C A T C C C G C A T C C C G C A T C C C G C A T C C C G C A T C C G C

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

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

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

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

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

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

SybGREEN qpcr Primers and Standards

SybGREEN qpcr Primers and Standards SybGREEN qpcr Primers and Standards Tools for RT-PCR Application Guide Table of Contents Package Contents and Storage Conditions... 1 Introduction... 3 SybGREEN qpcr Primer Pairs... 3 Gene Specific qpcr

More information

Reagents for PCR. Expertise in Amplification

Reagents for PCR. Expertise in Amplification Reagents for PCR Expertise in Amplification Selection Guide Hot Start Enzymes for Specific Requirements PCR Application: Routine PCR Fast PCR Hot Start Multiplex/ Sequencing Wide Variety/ Easy Access High

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

Gene Expression Assays

Gene Expression Assays APPLICATION NOTE TaqMan Gene Expression Assays A mpl i fic ationef ficienc yof TaqMan Gene Expression Assays Assays tested extensively for qpcr efficiency Key factors that affect efficiency Efficiency

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

Certificate of Analysis

Certificate of Analysis Certificate of Analysis SMART-Seq v4 3 DE Kit Components Catalog No(s). Amount Lot Number 635042 (Not sold separately; Sold as a part of 635040) 96 rxns Specified on product label. 635043 (Not sold separately;

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

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

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

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

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

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

Highly specific and sensitive quantitation

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

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

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

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

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

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

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

2. Convert total mrna to cdna with reverse transcriptase.

2. Convert total mrna to cdna with reverse transcriptase. Testing Gene Expression by Reverse Transcriptase PCR (rt-pcr) Overview Introduction: PCR is one method in molecular biology to examine the expression of mrna from a gene. The key element required to utilize

More information

Premium PCR Enzymes. High Fidelity PCR Real Time PCR RT- PCR qrt- PCR High-Speed PCR Long PCR Routine PCR Multiplex PCR Hot-Start PCR GC-Rich PCR

Premium PCR Enzymes. High Fidelity PCR Real Time PCR RT- PCR qrt- PCR High-Speed PCR Long PCR Routine PCR Multiplex PCR Hot-Start PCR GC-Rich PCR Premium PCR Enzymes Introducing SapphireAmp "Load'N'Go" fast PCR master mix... p 15 EmeraldAmp "Load'N'Go" PCR master mixes for superior performance on GC or AT rich regions...p 19 SYBR Premix Ex Taq (Tli

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

4 General PCR Methods Page

4 General PCR Methods Page Table of Contents General PCR Methods Page PCR Protocol Selection Guide...65.1 Basic PCR... 66.1.1 Hot Start PCR - The new Standard... 66.1.1.1 Reagents and Equipment Required... 66.1.1.2 General Considerations

More information

Real-time PCR: Understanding C t

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

More information

StrataScript One-Tube RT-PCR System with Easy-A High-Fidelity PCR Cloning Enzyme

StrataScript One-Tube RT-PCR System with Easy-A High-Fidelity PCR Cloning Enzyme StrataScript One-Tube RT-PCR System with Easy-A High-Fidelity PCR Cloning Enzyme INSTRUCTION MANUAL Catalog #600168 Revision #056001a For in Vitro Use Only *600168-12_056001a/* LIMITED PRODUCT WARRANTY

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

Factors Influencing Multiplex Real-Time 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].

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

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

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

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

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

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

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

Enzymes & Reagents. ...for PCR, Real Time PCR and RT-PCR

Enzymes & Reagents. ...for PCR, Real Time PCR and RT-PCR Enzymes & Reagents...for PCR, Real Time PCR and RT-PCR PCR DNA Polymerases EuroClone recombinant DNA Polymerases are produced and purified from E.coli. Severe and rigorous production procedures ensure

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

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

Technical Bulletin #176. Avoiding DNA Contamination in RT-PCR

Technical Bulletin #176. Avoiding DNA Contamination in RT-PCR 1 of 7 7/12/2007 8:27 PM Shop My Account View Cart nmlkji Products nmlkj Documents Technical Resources > Help Desk > Technical Bulletins Technical Bulletin #176 Avoiding DNA Contamination in RT-PCR A frequent

More information

Relative Quantification of mirna Target mrnas by Real-Time qpcr. 1 Introduction. Gene Expression Application Note No. 4

Relative Quantification of mirna Target mrnas by Real-Time qpcr. 1 Introduction. Gene Expression Application Note No. 4 Gene Expression Application Note No. 4 Relative Quantification of mirna Target mrnas by Real-Time qpcr Ute Ernst, Jitao David Zhang, Anja Irsigler, Stefan Wiemann, Ulrich Tschulena Division: Molecular

More information

ProtoScript Taq RT-PCR Kit

ProtoScript Taq RT-PCR Kit DNA AMPLIFICATION & PCR ProtoScript Taq RT-PCR Kit Instruction Manual NEB #E6400S Store at 20 C ProtoScript Taq RT-PCR Kit Table of Contents: Introduction... 2 Quality Controls.... 2 Important Factors

More information

REAL-TIME PCR: Put the odds in your favor with SuperScript RT. FROM THEORY TO PRACTICE

REAL-TIME PCR: Put the odds in your favor with SuperScript RT. FROM THEORY TO PRACTICE i REAL-TIME PCR: FROM THEORY TO PRACTICE ii Put the odds in your favor with SuperScript RT. Engineered to be RNase H and incredibly thermostable, SuperScript III RT delivers robust first-strand synthesis

More information

Thermo Scientific Advanced PCR Enzymes

Thermo Scientific Advanced PCR Enzymes Thermo Scientific Advanced PCR Enzymes Thermo Scientific Phusion High-Fidelity DNA Polymerase Thermo Scientific Phusion Hot Start II High-Fidelity DNA Polymerase Thermo Scientific Phire Hot Start II DNA

More information

Upgrade to the gold standard for high-fidelity PCR

Upgrade to the gold standard for high-fidelity PCR 3 Thermo Scientific Phusion and Phire DNA Polymerases Accurate, fast and powerful Polymerases for better PCR Extremely accurate, fast and robust amplification Instant activation hot start technology Direct

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

Upgrade to the gold standard for high-fidelity PCR

Upgrade to the gold standard for high-fidelity PCR 3 Thermo Scientific Phusion and Phire DNA s Accurate, fast and powerful s for better PCR Extremely accurate, fast and robust amplification Instant activation hot start technology Direct loading on gels

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

Taq98 Hot Start 2X Master Mix

Taq98 Hot Start 2X Master Mix Taq98 Hot Start 2X Master Mix Optimized for 98C Denaturation Lucigen Corporation 2905 Parmenter St, Middleton, WI 53562 USA Toll Free: (888) 575-9695 (608) 831-9011 FAX: (608) 831-9012 lucigen@lucigen.com

More information

PCR Reagents. The Highest Level of PCR Performance. PCR Enzymes RT-PCR Reagents QPCR and QRT-PCR Reagents dntps

PCR Reagents. The Highest Level of PCR Performance. PCR Enzymes RT-PCR Reagents QPCR and QRT-PCR Reagents dntps A m p l i f i c a t i o n PCR Reagents The Highest Level of PCR Performance PCR Enzymes RT-PCR Reagents QPCR and QRT-PCR Reagents dntps Stratagene Has Yo High-Fidelity Application General Cloning TA/UA

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

Recombinant DNA & Genetic Engineering. Tools for Genetic Manipulation

Recombinant DNA & Genetic Engineering. Tools for Genetic Manipulation Recombinant DNA & Genetic Engineering g Genetic Manipulation: Tools Kathleen Hill Associate Professor Department of Biology The University of Western Ontario Tools for Genetic Manipulation DNA, RNA, cdna

More information

POLYMERASES & AMPLIFICATION. OneTaq RT-PCR Kit. Instruction Manual. NEB #E5310S 30 reactions

POLYMERASES & AMPLIFICATION. OneTaq RT-PCR Kit. Instruction Manual. NEB #E5310S 30 reactions POLYMERASES & AMPLIFICATION OneTaq RT-PCR Kit Instruction Manual NEB #E5310S 30 reactions ISO 9001 Registered Quality Management ISO 14001 Registered Environmental Management ISO 13485 Registered Medical

More information

Optimizing buffer conditions and high-throughput screening of ligand protein binding

Optimizing buffer conditions and high-throughput screening of ligand protein binding APPLICATION NOTE Protein Thermal Shift technology Protein Thermal Shift technology Optimizing buffer conditions and high-throughput screening of ligand protein binding The Protein Thermal Shift assay measures

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

Global MicroRNA Amplification Kit

Global MicroRNA Amplification Kit Global MicroRNA Amplification Kit Store kit at -20 C on receipt (ver. 3-060901) A limited-use label license covers this product. By use of this product, you accept the terms and conditions outlined in

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

RIBOPROTECT. RNase Inhibitor RT33-020, RT33-100

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

More information

Competitive PCR Guide

Competitive PCR Guide Lit. # L0126 Rev. 8/99 Competitive PCR Guide Table of Contents I. What is Competitive PCR? A. Difficulties of Quantitative Analysis in Normal PCR... 2 B. Principle of Competitive PCR... 3 C. Competitive

More information

Table of Contents T ABLE OF C ONTENTS CHAPTER 1 CHAPTER 2 CHAPTER 3 CHAPTER 4 CHAPTER 5 CHAPTER 6 CHAPTER 7 CHAPTER 8 CHAPTER 9 CHAPTER 10 CHAPTER 11

Table of Contents T ABLE OF C ONTENTS CHAPTER 1 CHAPTER 2 CHAPTER 3 CHAPTER 4 CHAPTER 5 CHAPTER 6 CHAPTER 7 CHAPTER 8 CHAPTER 9 CHAPTER 10 CHAPTER 11 T ABLE OF C ONTENTS Table of Contents CHAPTER 1 CHAPTER 2 CHAPTER 3 CHAPTER 4 CHAPTER 5 CHAPTER 6 CHAPTER 7 CHAPTER 8 CHAPTER 9 CHAPTER 10 CHAPTER 11 CHAPTER 12 APPENDIX A The Basics of PCR and RT-PCR.................................................

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

PCR Enzyme Selection Guide

PCR Enzyme Selection Guide PCR Enzyme Selection Guide T: 1 (800) 313 7224, W: www.biobasic.com, E: order@biobasic.com, Copyright Bio Basic Inc. All rights reserved. Contents 1. PCR Enzymes Overview and Comparison. 2. Featured Product

More information

Overview. KAPAHiFi HotStart is the engineered KAPAHiFi DNA Polymerase with an antibody-based hot start technology and improved buffer system.

Overview. KAPAHiFi HotStart is the engineered KAPAHiFi DNA Polymerase with an antibody-based hot start technology and improved buffer system. Overview KAPAHiFi HotStart is the engineered KAPAHiFi DNA Polymerase with an antibody-based hot start technology and improved buffer system. KAPAHiFi HotStart exhibits: World leading fidelity confirmed

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

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

BioBank TM cdna Kit. Instructions for the use of BioBank TM cdna in real-time PCR. BB BioBank control cdna BioBank TM cdna Kit Instructions for the use of BioBank TM cdna in real-time PCR BB BioBank control cdna Contents Introduction 3 Kit Contents 4 Reagents and Equipment to Be Supplied by User 4 PrimerDesign

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

5PCR, real-time PCR, reverse transcription, and cloning

5PCR, real-time PCR, reverse transcription, and cloning 5PCR, real-time PCR, reverse transcription, and cloning 166 www.qiagen.com QIAGEN Product Guide 2007 PCR, real-time PCR, reverse transcription, and cloning 5 5.1 PCR and RT-PCR www.qiagen.com/pg/pcr Selection

More information

Řekněte si o vzorky zdarma!

Řekněte si o vzorky zdarma! strana 1 z 6 Ceník platí v souladu s Obchodními podmínkami LAB MARK od 15.10.2015 Core Reagents for Molecular Biology www.bioline.com Řekněte si o vzorky zdarma! PCR ENZYME & MIXES DNA Polymerases - For

More information

PrimeScript High Fidelity RT-PCR Kit

PrimeScript High Fidelity RT-PCR Kit For Research Use PrimeScript High Fidelity RT-PCR Kit Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Materials Required but not Provided... 4 IV. Storage... 4 V. Fidelity

More information

Dynabeads mrna DIRECT Micro Kit

Dynabeads mrna DIRECT Micro Kit USER GUIDE Dynabeads mrna DIRECT Micro Kit Catalog Number 61021 Revision 004 Revision Date 14 May 2012 For Research Use Only. Not for human or animal therapeutic or diagnostic use. For Research Use Only.

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

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

Application Guide... 2

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

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

PreciseTM Whitepaper

PreciseTM Whitepaper Precise TM Whitepaper Introduction LIMITATIONS OF EXISTING RNA-SEQ METHODS Correctly designed gene expression studies require large numbers of samples, accurate results and low analysis costs. Analysis

More information

Agilent SureCycler 8800 SURELY BETTER PCR SOLUTIONS CHOOSING THE RIGHT THERMAL CYCLER HAS NEVER BEEN EASIER

Agilent SureCycler 8800 SURELY BETTER PCR SOLUTIONS CHOOSING THE RIGHT THERMAL CYCLER HAS NEVER BEEN EASIER Agilent SureCycler 8800 SURELY BETTER PCR SOLUTIONS CHOOSING THE RIGHT THERMAL CYCLER HAS NEVER BEEN EASIER SURELY BETTER PCR FUNCTIONALITY FOR YOUR LAB Remote access Use a web-enabled device to monitor

More information