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:
Choose a product
Size
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:
選択製品の構成品内容
| 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
Search for SDS分析証明書
使用制限
For Laboratory Use. Outside of the United States, this product is intended for research use only unless otherwise stated.保存条件
選択製品の構成品内容
| 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 SDS分析証明書
使用制限
For Laboratory Use. Outside of the United States, this product is intended for research use only unless otherwise stated.保存条件
選択製品の構成品内容
| 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 |
選択製品の構成品内容
| 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 |
選択製品の構成品内容
| 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 |
選択製品の構成品内容
| 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 |
選択製品の構成品内容
| 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 |
選択製品の構成品内容
| 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 |
Resources
No related resources available
Related Products
類似製品
AMV Reverse Transcriptase
高い反応温度でも安定な逆転写酵素
M5101, M5108, M9004
M-MLV Reverse Transcriptase
cDNA 合成のための RNA 依存性 DNA ポリメラーゼ
M1701, M1705, M5313
M-MLV Reverse Transcriptase, RNase H Minus
完全長 cDNA の割合の高い長鎖 cDNA およびライブラリー調製に有用
M5301
M-MLV Reverse Transcriptase, RNase H Minus, Point Mutant
長鎖 鋳型 RNA (<5kb) からの cDNA 合成に適した酵素
M3681, M3682, M3683
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).
