SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase
SuperScript&trade; III One-Step RT-PCR System with Platinum&trade; <i>Taq</i> DNA Polymerase
Invitrogen™

SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase

The SuperScript III One-Step RT-PCR System with Platinum Taq DNA Polymerase is designed for the sensitive, reproducible, end-point detection andRead more
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Catalog NumberNo. of Reactions
12574026100 Reactions
1257401825 Reactions
Catalog number 12574026
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1.026,65
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1.122,00
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No. of Reactions:
100 Reactions
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Price (EUR)
1.026,65
Online Exclusive
1.122,00
Save 95,35 (8%)
Each
Add to cart

The SuperScript III One-Step RT-PCR System with Platinum Taq DNA Polymerase is designed for the sensitive, reproducible, end-point detection and analysis of RNA molecules by RT-PCR. Using this convenient one-step formulation, you can perform both cDNA synthesis and PCR amplification in a single tube using gene-specific primers and target RNAs from either total RNA or mRNA. The system uses a mixture of SuperScript III Reverse Transcriptase and Platinum Taq DNA polymerase in an optimized reaction buffer, and it can detect a wide range of RNA targets, from 200 bp to 4.5 kb. The amount of starting material can range from 0.01 pg to 1 μg of total RNA.

Key advantages of SuperScript III One-Step RT-PCR System with Platinum Taq DNA Polymerase
• Sensitive detection down to 0.01 pg total RNA
• Convenient one-step format for increased speed, convenience, and less reaction-to-reaction variability
• SuperScript III RT for cDNA synthesis up to 55°C, for more specific priming with gene specific primers
• Amplicon size—compatible detection of targets up to 4.5 kb in length for greater flexibility

SuperScript III Reverse Transcriptase
SuperScript III Reverse Transcriptase is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme can synthesize cDNA at a temperature range of 45–60°C, providing increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases. Because SuperScript III RT is not significantly inhibited by ribosomal and transfer RNA, it can be used to synthesize cDNA from total RNA.

Platinum Taq DNA Polymerase
Platinum Taq DNA Polymerase is recombinant Taq DNA polymerase complexed with a proprietary antibody that blocks polymerase activity at ambient temperatures. Activity is restored after the denaturation step in PCR cycling at 94°C, providing an automatic 'hot start' in PCR for increased sensitivity, specificity, and yield.

Note: This kit has been optimized for end-point RT-PCR. For RT–qPCR in this product line, use the SuperScript III Platinum One–Step Quantitative RT-PCR System.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Final Product TypePCR Amplified cDNA
FormatOne-Step RT-PCR System
Hot StartBuilt-In Hot Start
No. of Reactions100 Reactions
Optimal Reaction Temperature50°C
PolymerasePlatinum Taq DNA Polymerase
Quantity100 rxns
Reaction FormatPremixed Components
Reagent TypeReverse Transcription
Reverse TranscriptaseSuperScript III
Ribonuclease H ActivityReduced
Shipping ConditionDry Ice
Size (Final Product)Up to 4.5 kb
Starting MaterialRNA
Technique1-Step RT-PCR
Detection MethodGel Electrophoresis
GC-Rich PCR PerformanceHigh
PCR Method1-step RT-PCR
Reaction Speed30 min.
Unit SizeEach
Contents & Storage

• SuperScript III RT/Platinum Taq Mix (200 μL)
• 2X Reaction Mix (3 x 1 mL)
• 5 mM Magnesium Sulfate (500 μL)

Store at –10 to –30°C.

Frequently asked questions (FAQs)

How can I remove genomic DNA contamination from my sample prior to performing RT-PCR?

We recommend using ezDNase (Cat. No. 11766051). ezDNase Enzyme's high specificity for double-stranded DNA enables efficient and fast genomic DNA removal without reduction in the quality or quantity of RNA. ezDNase Enzyme is heat-labile and so can be easily deactivated by heat treatment at moderate temperature (55 degrees C). These features make ezDNase Enzyme an excellent choice for genomic DNA removal prior to reverse transcription reactions.

How much RNA should be employed for first-strand cDNA synthesis?

The amount of RNA template for a cDNA synthesis is highly flexible and depends upon the amount of sample available and an individual's need. In general, 1 µg total RNA is used in a typical 20-µL RT reaction.

Find additional tips, troubleshooting help, and resources within ourReverse Transcription and RACE Support Center.

Should I treat the cDNA with RNase H prior to downstream processing?

RNase H treatment is not always necessary. Many PCR reactions work without it. However, for cDNA synthesized with RNase H-deficient reverse transcriptases (like SuperScript II, III, and IV), RNA/cDNA hybrids—especially GC-rich ones—may not denature well, reducing PCR sensitivity. RNase H treatment can help in such cases. Additionally, RNase H treatment is beneficial for cloning larger fragments.

What percentage of RNA is converted to cDNA when performing reverse transcription?

This depends highly on the quality of the sample. mRNA itself makes up 1-5% of total RNA. Depending on the primer and enzyme used, reverse transcription can covert >70% of that into cDNA.

Find additional tips, troubleshooting help, and resources within our Reverse Transcription and RACE Support Center.

I'm setting up my RT reaction and am trying to decide whether I should use random primers, oligo(dT) primer, gene-specific primer, or oligo(dT)/random mix primers. What would you suggest?

Random primers are the best choice for degraded RNA, RNA with heavy secondary structure, non-polyadenylated RNA, or prokaryotic RNA. It is recommended only for two-step RT-PCR, and typically gives the highest yields, although the cDNA may not necessarily be full length. Oligo(dT) primers are good to use when trying to recover full-length cDNA from 2-step RT-PCR. The reaction is influenced by secondary structure and RNA quality. Gene specific primers should be used for very specific, mainly one-step RT-PCR reactions.

Find additional tips, troubleshooting help, and resources within our Reverse Transcription and RACE Support Center.

Citations & References (2)

Citations & References
Abstract
Clinical accuracy of a PLEX-ID flu device for simultaneous detection and identification of influenza viruses A and B.
Authors:Tang YW, Lowery KS, Valsamakis A, Schaefer VC, Chappell JD, White-Abell J, Quinn CD, Li H, Washington CA, Cromwell J, Giamanco CM, Forman M, Holden J, Rothman RE, Parker ML, Ortenberg EV, Zhang L, Lin YL, Gaydos CA,
Journal:J Clin Microbiol
PubMed ID:23077123
Respiratory tract infections caused by influenza A and B viruses often present nonspecifically, and a rapid, high-throughput laboratory technique that can identify influenza viruses is clinically and epidemiologically desirable. The PLEX-ID Flu assay (Abbott Molecular Inc., Des Plaines, IL) incorporates multilocus PCR and electrospray ionization-mass spectrometry to detect and differentiate ... More
Two isoforms of Npap60 (Nup50) differentially regulate nuclear protein import.
Authors:Ogawa Y, Miyamoto Y, Asally M, Oka M, Yasuda Y, Yoneda Y,
Journal:Mol Biol Cell
PubMed ID:20016008
Npap60 (Nup50) is a nucleoporin that binds directly to importin alpha. In humans, there are two Npap60 isoforms: the long (Npap60L) and short (Npap60S) forms. In this study, we provide both in vitro and in vivo evidence that Npap60L and Npap60S function differently in nuclear protein import. In vitro binding ... More