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
Catalog Number:
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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
What is GoScript™ Reverse Transcriptase?
GoScript™ Reverse Transcriptase is a recombinant M-MLV enzyme and proprietary reaction buffer combination optimized for first-strand cDNA synthesis in RT-qPCR workflows. GoScript™ Reverse Transcriptase is engineered for robust cDNA production from rare and abundant targets across a wide range of sizes, even in the presence of reverse transcriptase inhibitors.
What are the Benefits of GoScript™ Reverse Transcriptase?
- Processivity: Reverse transcribes long messages and templates, including those with significant secondary structure
- Sensitivity: Reverse transcribes rare transcripts alongside abundant targets in the same reaction
- Enzyme efficiency: High specific activity reduces the amount of enzyme required per reaction, lowering cost without sacrificing yield
Which GoScript™ Format is Right For My Experiment?
| GoScript™ Reverse Transcription System (Complete Kit) | GoScript™ Reverse Transcription Mix + Oligo(dT) | GoScript™ Reverse Transcription Mix + Random Primers | GoScript™ Reverse Transcriptase (Enzyme) | |
|---|---|---|---|---|
| Best Use | Gene expression from total RNA, mRNA or synthetic RNA | mRNA / poly(A)+ RNA targets | All RNA types including non-polyadenylated & prokaryotic | Custom reactions with your own primers & accessories |
| GoScript™ Reverse Transcriptase | ✔ | ✔ | ✔ | ✔ |
| MgCl₂ | ✔ | ✔ | ✔ | ✔ |
| dNTP Mix | ✔ | ✔ | ✔ | – |
| Recombinant RNasin® Inhibitor | ✔ | ✔ | ✔ | – |
| Random Primers | ✔ | – | ✔ | – |
| Oligo(dT)₁₅ Primer | ✔ | ✔ | – | – |
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
Quick Protocols
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size | 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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Use Restrictions
For Laboratory Use. Outside of the United States, this product is intended for research use only unless otherwise stated.Storage Conditions
What's in the box?
| Item | Part # | Size | 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 SDSCertificate of Analysis
Use Restrictions
For Laboratory Use. Outside of the United States, this product is intended for research use only unless otherwise stated.Storage Conditions
What's in the box?
| Item | Part # | Size | 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 SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
What's in the box?
| Item | Part # | Size | 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 SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
What's in the box?
| Item | Part # | Size | 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 SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
What's in the box?
| Item | Part # | Size | 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 SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
What's in the box?
| Item | Part # | Size | 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 SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
What's in the box?
| Item | Part # | Size | 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 SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
Resources
No related resources available
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Frequently Used With
ReliaPrep™ RNA Miniprep Systems
Simple, 30-minute protocol to extract total RNA from as few as 100 cells or 0.25mg of tissue.
Z6014, Z6015, Z6010, Z6011, Z6012, Z6110, Z6111, Z6112
GoTaq® Hot Start Polymerase
Native Taq DNA polymerase with room-temperature setup for hot-start PCR.
M5001, M5005, M5006, M5008
Frequently Asked Questions
What primer types work with GoScript™ Reverse Transcriptase?
GoScript™ Reverse Transcriptase is compatible with Oligo(dT)₁₅ primers (for poly-A-tailed mRNA), random primers (for total RNA or rRNA targets), and gene-specific primers. For most RT-qPCR applications, Oligo(dT)₁₅ or random primers are recommended. The GoScript™ Reverse Transcription System includes both primer types.
Can GoScript™ reverse transcribe RNA with significant secondary structure or inhibitors?
Yes. GoScript™ Reverse Transcriptase is formulated to maintain enzyme activity in the presence of common RT-qPCR inhibitors, such as ethanol. GoScript™ Reverse Transcriptase efficiently synthesizes cDNA from highly structured targets.
What is the difference between the GoScript™ Reverse Transcription System and the GoScript™ Reverse Transcriptase enzyme?
The GoScript™ Reverse Transcription System (Cat.# A5001) is a complete kit containing the enzyme, 5X Reaction Buffer, MgCl₂, primers (Oligo(dT)₁₅ and random), PCR Nucleotide Mix, Recombinant RNasin® Ribonuclease Inhibitor, and Nuclease-Free Water. The standalone GoScript™ Reverse Transcriptase (Cat.# A5003) includes only the enzyme, 5X Reaction Buffer, and MgCl₂ for labs that supply their own accessories.
Is GoScript™ Reverse Transcriptase compatible with one-step RT-qPCR?
GoScript™ Reverse Transcriptase is validated for two-step RT-qPCR. For one-step RT-qPCR, see the GoTaq® One-Step RT-qPCR System (Cat.# A6020) or the GoTaq® Probe One-Step RT-qPCR System (Cat.# A6120).
