Certificate of Analysis

GoScript™ Reverse Transcriptase

Part# 9PIA500
Cat.# Size
A5002 10 reactions
A5003 100 reactions Revised 10/16
A5004 500 reactions

Description: GoScript™ Reverse Transcriptase(a) utilizes M-MLV and state-of-the-art buffer technology designed for qPCR
to deliver robust, reliable cDNA synthesis of a full range of rare and abundant transcripts, even in the presence of inhibitors.
GoScript™ Reverse Transcriptase(a) is qualified for use in qPCR, including GoTaq® qPCR and Plexor® qPCR systems for
performing RT-qPCR.
GoScript™ Reverse Transcriptase is provided in quantities sufficient for 10–500 first-strand cDNA synthesis reactions of
*AF9PIA500 1016A500*
AF9PIA500 1016A500
20µl each. Additional components are required to perform the reactions. Contains one of the following:

Cat.# A5002
Part No. Component Size
A501A GoScript™ Reverse Transcriptase 10µl
A500A GoScript™ 5X Reaction Buffer 100µl
A351B MgCl2, 25mM 750µl

Cat.# A5003
Part No. Component Size
A501C GoScript™ Reverse Transcriptase 100µl
A500C GoScript™ 5X Reaction Buffer 600µl
A351H MgCl2, 25mM 1.2ml

Cat.# A5004 Promega Corporation

2800 Woods Hollow Road
Part No. Component Size Madison, WI 53711-5399 USA
A501D GoScript™ Reverse Transcriptase 500µl Telephone 608-274-4330
A500D GoScript™ 5X Reaction Buffer 2 × 1.25ml Toll Free 800-356-9526
A351H MgCl2, 25mM 3 × 1.2ml Fax 608-277-2516
Internet www.promega.com
Concentration: GoScript™ Reverse Transcriptase is supplied at a concentration of 160u/µl.
Storage Conditions: Store at –20°C.
Expiration Date: See the product label for the expiration date. PRODUCT USE LIMITATIONS, WARRANTY DISCLAIMER
Biological Source: Recombinant E. coli strain. Promega manufactures products for a number of
intended uses. Please refer to the product label for the
intended use statements for specific products.
Promega products contain chemicals which may be
harmful if misused. Due care should be exercised with
all Promega products to prevent direct human contact.
Quality Control Assays Each Promega product is shipped with documentation
stating specifications and other technical information.
Promega products are warranted to meet or exceed the
stated specifications. Promega's sole obligation and the
This lot passes the following quality control specifications: customer's sole remedy is limited to replacement of
products free of charge in the event products fail to
Unit Definition: One unit is defined as the amount of enzyme required to catalyze the transfer of 1nmol of deoxynucleotide perform as warranted. Promega makes no other war-
ranty of any kind whatsoever, and SPECIFICALLY DIS-
into acid-precipitable material in 10 minutes at 37°C. CLAIMS AND EXCLUDES ALL OTHER WARRANTIES
OF ANY KIND OR NATURE WHATSOEVER, DIRECTLY
First-Strand cDNA Synthesis: First-strand cDNA is synthesized using 1.25µl of GoScript™ enzyme, 1µg of a 6.5kb con- OR INDIRECTLY, EXPRESS OR IMPLIED, INCLUDING,
WITHOUT LIMITATION, AS TO THE SUITABILITY,
trol RNA, oligo(dT) primer and radiolabeled dNTP. The specification is the conversion of >12% of mRNA to cDNA. Full- PRODUCTIVITY, DURABILITY, FITNESS FOR A PAR-
length cDNA is observed by gel electrophoresis and autoradiography. TICULAR PURPOSE OR USE, MERCHANTABILITY,
CONDITION, OR ANY OTHER MATTER WITH
RESPECT TO PROMEGA PRODUCTS. In no event shall
Promega be liable for claims for any other damages,
whether direct, incidental, foreseeable, consequential,
Amplification: or special (including but not limited to loss of use, rev-
enue or profit), whether based upon warranty, contract,
RT-PCR: When approximately 100 copies of a 1.2kb Kanamycin Positive Control RNA (Cat.# C1381) are reverse tran- tort (including negligence) or strict liability arising in
connection with the sale or the failure of Promega prod-
scribed at 42°C and amplified, the result is a clear, discrete 323bp DNA product as visualized on an agarose gel by ethidium ucts to perform in accordance with the stated specifica-
bromide staining. tions.

RT-qPCR: One microgram of total RNA positive control (Part# C199A) is reverse transcribed at 42°C for 1 hour. The result-
ing cDNA is serially diluted over six orders of magnitude and amplified using GoTaq® qPCR Master Mix, 2X (Part# A600A),
to generate a standard curve. The standard curve has an R2 ≥ 0.990. © 2009–2012, 2016 Promega Corporation. All Rights
Reserved.
GoTaq and Plexor are registered trademarks of
Promega Corporation. GoScript is a trademark of
Promega Corporation.
Products may be covered by pending or issued
patents or may have certain limitations. Please visit
our Web site for more information.
All prices and specifications are subject to change
without prior notice.
Product claims are subject to change. Please contact
Promega Technical Services or access the Promega
online catalog for the most up-to-date information on
Promega products.
Signed by: Part# 9PIA500
R. Wheeler, Quality Assurance Printed in USA, Revised 10/16.
(a)U.S.
Pat. No. 6,242,235, Australian Pat. No. 761757, Canadian Pat. No. 2,335,153, Chinese Pat. No. ZL99808861.7, Hong Kong
Pat. No. HK 1040262, Japanese Pat. No. 3673175, European Pat. No. 1088060 and other patents pending.
 Usage Information
I. First-Strand cDNA Synthesis II. cDNA Quantification Using qPCR
The following procedure can be used to convert up to 5µg of total RNA or up to 500ng of cDNA synthesized using GoScript™ Reverse Transcriptase can be amplified and quantified
using the GoTaq® qPCR Master Mix or the Plexor® qPCR Systems. cDNA samples may be used
poly(A) RNA into first-strand cDNA.
directly or diluted prior to amplification. As a starting point for dilution, dilute sample and refer-
1. Mix and briefly centrifuge each component before use. Combine the following: ence standard cDNA reactions 1:10, then add 5µl of these diluted reactions to the reaction mix.
For additional information, refer to the GoTaq® qPCR Master Mix Technical Manual, #TM318,
Component Volume
or to the Plexor® qPCR System Technical Manual, #TM262.
Experimental RNA (up to 5µg/reaction) Xµl
Primer [Oligo(dT)15 (0.5µg/reaction) and/or 1. Heat inactivate the reverse transcription reaction.
Random Primer (0.5µg/reaction) or
2. Proceed directly to GoTaq® or Plexor® qPCR target-specific quantitative analysis of the
gene-specific primer (10–20pmol/reaction)] Xµl
cDNA.
Nuclease-Free Water Xµl
Final volume 5µl 3. Alternatively, heat-inactivated reactions may be stored frozen for future use.
2. Heat in a 70°C heat block for 5 minutes. Immediately chill in ice water for at least 4. GoTaq® and Plexor® qPCR accommodate the addition of up to 20% of the total reaction
5 minutes. Centrifuge 10 seconds in a microcentrifuge. Store on ice until reverse volume as template GoScript™ Reverse Transcriptase reaction volume.
transcription mix is added.
5. The cDNA may be added directly to the qPCR, as undiluted reverse transcription reaction
3. Prepare the reverse transcription reaction mix, 15µl for each cDNA reaction. Combine product, or it may be diluted.
on ice, in the order listed.
6. The dilution factor must be experimentally determined to be appropriate for the amount of
Component Volume RNA template mass and proportional reverse-transcript representation in the cDNA sam-
GoScript™ 5X Reaction Buffer 4.0µl ple.
MgCl2 (final concentration 1.5–5.0mM)1 1.2–6.4µl 7. Generally, for cDNA quantitation, using the default analysis settings of many real-time
PCR Nucleotide Mix (final concentration 0.5mM each dNTP)2 1.0µl instruments, the amount of cDNA used as template in GoTaq® qPCR should not exceed
Recombinant RNasin® Ribonuclease Inhibitor (optional) 20units that correlating with a proportional 100ng of input total RNA. The cDNA generated from
GoScript™ Reverse Transcriptase 1.0µl highly abundant transcripts can be detected in less than 1pg of total RNA.
Nuclease-Free Water (to a final volume of 15µl) Xµl
Final volume 15µl
1Mg2+ concentration should be optimized to 1.5–5.0mM (MgCl2 provided at 25mM).
2Ifisotopic or nonisotopic incorporation is desired for monitoring first-strand cDNA syn-
thesis, α[32P]-dCTP or other modified nucleotides may be supplemented into the PCR
Nucleotide Mix. See Section 4.D of TM316, for analysis suggestions.

4. Combine 15µl of reverse transcription mix with 5µl of RNA and primer mix.

5. Anneal in a heat block at 25°C for 5 minutes.

6. Extend in a heat block at 42°C for up to one hour.

Reactions can be stopped at this point for analysis of the cDNA or may be frozen for
long-term storage.

7. Inactivate Reverse Transcriptase: Before proceeding with qPCR, inactivate the reverse
transcriptase in a heat block at 70°C for 15 minutes.

Part# 9PIA500
Printed in USA. Revised 10/16.

Promega Corporation · 2800 Woods Hollow Road·Madison, WI 53711-5399 U.S.A. · Toll Free in the USA 800-356-9526 · Telephone 608-274-4330 · Internet www.promega.com