GoScript™ Reverse Transcriptase

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 Transcriptase is available in convenient mixes with either Oligo (dT) primers or random primers, as part of a complete kit, and as a stand-alone enzyme.

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

efficient cDNA Synthesis Over a Range of Input RNA 13870MB-W — 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

GoScript reverse transcriptase resistance to inhibitors 8675MB-W — 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 reverse transcriptase 14113MA-W

GoScript™ RT Mix, Random Primers, TBP Variable RNA Input

GoScript reverse transcriptase 14114MA-W

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 dynamic range 8565MB-W-a

Goscript reverse transcriptase titration over 9 logs 8565MB-W-b

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

GoScript™ Reverse Transcriptase Product InformationPDF (109 KB) – English

GoScript™ Reverse Transcription System Technical ManualPDF (8371 KB) – English

GoScript™ Reverse Transcription Mix, Oligo(dT)PDF (120 KB) – English

GoScript™ Reverse Transcription Mix, Random PrimersPDF (120 KB) – English

Quick Protocols

GoScript™ Reverse Transcription System Quick Protocol FB114PDF (106 KB) – English

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
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
GoScript™ Reverse Transcriptase	A501D	1 × 500μl

SDS

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Certificate of Analysis

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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	A351B	1 × 750μl	25mM	View Product
GoScript™ 5X Reaction Buffer	A500B	1 × 300μl
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

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Certificate of Analysis

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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
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 SDS

Certificate of Analysis

Search by lot number

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 SDS

Certificate of Analysis

Search by lot number

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 SDS

Certificate of Analysis

Search by lot number

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 SDS

Certificate of Analysis

Search by lot number

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 SDS

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

Resources

Citations

Frequently Used With

ReliaPrep™ RNA Miniprep Systems

Simple, 30-minute protocol to extract total RNA from as few as 100 cells or 0.25mg tissue.

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

Reliable cDNA Synthesis for Sensitive Detection of Expression

Sensitive Reverse Transcriptase Ideal for Challenging Templates

One Reverse Transcriptase, Four Convenient Formats

Reverse Transcriptase Performance Data

Efficient cDNA Synthesis Over a Wide Range of Input RNA

Resistance to Common Inhibitors

Efficient Detection of Low-Copy Targets

GoScript™ RT Mix Oligo(dT), TBP Variable RNA Input

GoScript™ RT Mix, Random Primers, TBP Variable RNA Input

Sensitivity and Specificity

Protocols

Complete Protocol

Quick Protocols

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Resources

Citations

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Frequently Used With

ReliaPrep™ RNA Miniprep Systems

GoTaq® Hot Start Polymerase

Americas

Pacific Asia

Europe