GoScript™ Reverse Transcriptase
Reliable cDNA Synthesis for Sensitive Detection of Expression
- Available as a standalone enzyme, a complete reverse transcription kit or as a master mix with Oligo(dT) or Random Primers
- Sensitive transcription of both high-copy and low-copy messages
- Transcribes short and long transcripts; processes through secondary structure
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Catalog Number: A5003
Catalog Number: A5004
Catalog Number: A5000
Catalog Number: A5001
Catalog Number: A2790
Catalog Number: A2791
Catalog Number: A2800
Catalog Number: A2801
Sensitive Reverse Transcriptase Ideal for Challenging Templates
GoScript™ Reverse Transcriptase utilizes M-MLV reverse transcriptase enzyme and state-of-the-art buffer to drive robust, reliable cDNA synthesis of a full range of rare and abundant transcripts, even with difficult templates and in the presence of PCR inhibitors.
One Reverse Transcriptase, Four Convenient Formats
GoScript™ Reverse Transcription System
Includes GoScript™ Reverse Transcriptase, Reaction Buffer, MgCl2, dNTP Mix, Oligo(dT), Random Primers and Recombinant RNasin® Ribonuclease Inhibitor, and is designed for efficient synthesis of first-strand cDNA. The system components can effectively reverse transcribe RNA templates starting with total RNA, poly(A)+ mRNA or synthetic transcript RNA.
GoScript™ Reverse Transcription Mix, Oligo(dT)
GoScript™ Reverse Transcription Mix, Oligo(dT) includes an optimized set of reagents designed for efficient synthesis of first-strand cDNA. The Oligo(dT) priming initiates first-strand synthesis by annealing to the 3’ end of any polyadenylated RNA molecule. Reverse Transcription of total RNA or mRNA will yield oligo(dT)-primed products from any poly(A)+RNA. The population of cDNAs produced using this method allows many different downstream analyses from a single reverse transcription reaction.
GoScript™ Reverse Transcription Mix, Random Primers
Includes random primers for synthesis of first-strand cDNA in preparation for qPCR amplification. Priming using random primers is the most generally used method of initiating cDNA synthesis from a variety of RNA templates. Random primers can be used to prime first-strand cDNA synthesis from all RNA molecules, including those that do not possess a poly(A)+ tail and RNA isolated from prokaryotic sources.
GoScript™ Reverse Transcriptase
Includes GoScript™ M-MLV reverse transcriptase enzyme and 5X GoScript™ Reaction Buffer.
Reverse Transcriptase Performance Data
The data presented below illustrate the sensitivity, inhibitor resistance, and performance of GoScript™ Reverse transcriptase in cDNA Synthesis and RT-qPCR.
Efficient cDNA Synthesis Over a Wide Range of Input RNA
Direct quantification of six different targets in human total RNA using GoTaq® 2-Step RT-qPCR System. Nine parallel large-scale GoScript™ RT reactions were performed, containing log-fold variations of input human reference total RNA (10ng/µl to 0.1fg/µl). 10µl fractions of each reverse transcription reaction product, representing 100ng to 1fg of input RNA, were sampled directly into triplicate GoTaq® qPCR reactions, without dilution, for six separate monoplex assays. Each assay was designed to be specific for a different RNA target with an anticipated, proportionally different expression level: extremely high (5.8S rRNA), high (ACTB and B2M), moderate (HPRT ) or low (RPL29 and TBP1 ). In each different model, the results demonstrate the desired ≅3.3 cycle change per Δlog input, but with different Cqs reflecting relative proportion within the total RNA sample.
Resistance to Common Inhibitors
An in vitro-transcribed RNA was reverse transcribed using oligo(dT) primer according to the manufacturer’s instructions. Reverse transcription was carried out in the absence and presence of ethanol (5%, 10% and 20%). cDNA was analyzed by qPCR using GoTaq® qPCR Master Mix. Changes in cycle threshold values in the absence and presence of ethanol were determined and converted to relative change in transcript level = (2Cqinhibitor – Cqnone). Data are the average of three separate experiments.
Efficient Detection of Low-Copy Targets
Here, a series of 10 fold dilutions (1µg - 100pg) of total human RNA (Agilent, Cat #750500) was reverse transcribed using the GoScript™ Reverse Transcription Mix, Random Primers or Oligo(dT). The resulting cDNA was evaluated by qPCR amplification of the low-copy genes, APC or TBP, using the GoTaq® qPCR Master Mix (Cat. # A6002). Standard curves show that the primers had a calculated efficiency of 99% (APC), and 97% (TBP). Both values are within the range recommended by the MIQE guidelines. No-reverse transcriptase and no-template controls gave negative results.
GoScript™ RT Mix Oligo(dT), TBP Variable RNA Input
GoScript™ RT Mix, Random Primers, TBP Variable RNA Input
Sensitivity and Specificity
GoScript™ Reverse Transcriptase used in combination with GoTaq® qPCR Master Mix can provide detection of input templates over a 9-log order dynamic range.
GoScript™ Reverse Transcriptase cDNA synthesis coupled with GoTaq® qPCR Master Mix can detect input template over a 9-log order dynamic range. Panel A. Ten RNA template mixes were assembled to model a series of tenfold differences of a specific RNA abundance (1 × 102 to 1 × 1011 copies) in a constant mass of 10ng of human total RNA. cDNAs for each RNA sample were prepared using GoScript™ Reverse Transcriptase and amplified using GoTaq® qPCR at each level of total cDNA (n=3 each point). Inset (Panel A) shows dissociation profile for the 50-, 500- and 5,000-copy standard and 5-copy detected levels of MS2 + cDNA and NTC reactions, demonstrating specificity. Standard curve shown in Panel B illustrates the quantitation and detection of the “unknown” samples. ■ = tenfold standard dilutions; x = detection of 5-copy samples.
Protocols
Complete Protocol
Protocolos rápidos
Specifications
Catalog Number:
What's in the box?
| Item | Código | Tamanho | Concentration | Available Separately |
|---|---|---|---|---|
|
MgCl2 |
A351H | 1 × 1.2ml | 25mM | View Product |
|
GoScript™ 5X Reaction Buffer |
A500C | 1 × 600μl | View Product | |
|
GoScript™ Reverse Transcriptase |
A501C | 1 × 100μl |
SDS
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Restrições de uso
For Laboratory Use. Outside of the United States, this product is intended for research use only unless otherwise stated.Condições de armazenamento
What's in the box?
| Item | Código | Tamanho | Concentration | Available Separately |
|---|---|---|---|---|
|
MgCl2 |
A351H | 3 × 1.2ml | 25mM | View Product |
|
GoScript™ 5X Reaction Buffer |
A500D | 2 × 1.25ml | View Product | |
|
GoScript™ Reverse Transcriptase |
A501D | 1 × 500μl |
SDS
Search for SDSCertificado de Análise
Restrições de uso
For Laboratory Use. Outside of the United States, this product is intended for research use only unless otherwise stated.Condições de armazenamento
What's in the box?
| Item | Código | Tamanho | Concentration | Available Separately |
|---|---|---|---|---|
|
MgCl2 |
A351B | 1 × 750μl | 25mM | View Product |
|
GoScript™ 5X Reaction Buffer |
A500B | 1 × 300μl | View Product | |
|
GoScript™ Reverse Transcriptase |
A501B | 1 × 50μl | ||
|
Oligo(dT)15 Primer |
C110B | 1 × 50μg | 500μg/ml | View Product |
|
PCR Nucleotide Mix |
C114G | 1 × 200μl | 10mM | View Product |
|
Random Primers |
C118B | 1 × 50μg | View Product | |
|
Recombinant RNasin® Ribonuclease Inhibitor |
N251A | 1 × 2,500u | 40u/μl | View Product |
|
Nuclease-Free Water |
P119A | 1 × 1,250μl | View Product |
SDS
Search for SDSCertificado de Análise
Restrições de uso
For Research Use Only. Not for Use in Diagnostic Procedures.Condições de armazenamento
What's in the box?
| Item | Código | Tamanho | Concentration | Available Separately |
|---|---|---|---|---|
|
MgCl2 |
A351H | 1 × 1.2ml | 25mM | View Product |
|
GoScript™ 5X Reaction Buffer |
A500C | 1 × 600μl | View Product | |
|
GoScript™ Reverse Transcriptase |
A501C | 1 × 100μl | ||
|
Oligo(dT)15 Primer |
C110B | 1 × 50μg | 500μg/ml | View Product |
|
PCR Nucleotide Mix |
C114G | 1 × 200μl | 10mM | View Product |
|
Random Primers |
C118B | 1 × 50μg | View Product | |
|
Recombinant RNasin® Ribonuclease Inhibitor |
N251A | 1 × 2,500u | 40u/μl | View Product |
|
Nuclease-Free Water |
P119A | 2 × 1,250μl | View Product |
SDS
Search for SDSCertificado de Análise
Restrições de uso
For Research Use Only. Not for Use in Diagnostic Procedures.Condições de armazenamento
What's in the box?
| Item | Código | Tamanho | Available Separately |
|---|---|---|---|
GoScript™ Enzyme Mix |
A276A | 1 × 100μl | |
GoScript™ Reaction Buffer, Oligo(dT) |
A277A | 1 × 200μl | |
|
Nuclease-Free Water |
P119A | 1 × 1,250μl | View Product |
SDS
Search for SDSCertificado de Análise
Restrições de uso
For Research Use Only. Not for Use in Diagnostic Procedures.Condições de armazenamento
What's in the box?
| Item | Código | Tamanho | Available Separately |
|---|---|---|---|
GoScript™ Enzyme Mix |
A276A | 2 × 100μl | |
GoScript™ Reaction Buffer, Oligo(dT) |
A277A | 2 × 200μl | |
|
Nuclease-Free Water |
P119A | 2 × 1,250μl | View Product |
SDS
Search for SDSCertificado de Análise
Restrições de uso
For Research Use Only. Not for Use in Diagnostic Procedures.Condições de armazenamento
What's in the box?
| Item | Código | Tamanho | Available Separately |
|---|---|---|---|
GoScript™ Enzyme Mix |
A276A | 1 × 100μl | |
GoScript™ Reaction Buffer, Random Primers |
A278A | 1 × 200μl | |
|
Nuclease-Free Water |
P119A | 1 × 1,250μl | View Product |
SDS
Search for SDSCertificado de Análise
Restrições de uso
For Research Use Only. Not for Use in Diagnostic Procedures.Condições de armazenamento
What's in the box?
| Item | Código | Tamanho | Available Separately |
|---|---|---|---|
GoScript™ Enzyme Mix |
A276A | 2 × 100μl | |
GoScript™ Reaction Buffer, Random Primers |
A278A | 2 × 200μl | |
|
Nuclease-Free Water |
P119A | 2 × 1,250μl | View Product |
SDS
Search for SDSCertificado de Análise
Restrições de uso
For Research Use Only. Not for Use in Diagnostic Procedures.Condições de armazenamento
Resources
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