SuperScript™ III One-Step RT-PCR System with Platinum™ Taq High Fidelity DNA Polymerase
SuperScript&trade; III One-Step RT-PCR System with Platinum&trade; <i>Taq</i> High Fidelity DNA Polymerase
Citations & References (2)
Invitrogen™

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

The SuperScript III One-Step RT-PCR System with Platinum Taq High Fidelity is designed for convenient end-point detection and analysis ofRead more
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Catalog NumberNo. of Reactions
12574035100 Reactions
Catalog number 12574035
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1.012,65
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1.266,00
Save 253,35 (20%)
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No. of Reactions:
100 Reactions
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Price (EUR)
1.012,65
Online Exclusive
1.266,00
Save 253,35 (20%)
Each
Add to cart

The SuperScript III One-Step RT-PCR System with Platinum Taq High Fidelity is designed for convenient end-point detection and analysis of RNA molecules by one-step RT-PCR. This one-step formulation allows for both cDNA synthesis and PCR amplification to be performed in a single tube using gene-specific primers and target RNAs from either total RNA or mRNA. It also allows for the detection of a wide range of RNA targets (up to 10 kb in length) at variable concentrations (1 pg to 1 μg of total RNA). This system consists of two major components: SuperScript III RT/Platinum Taq High Fidelity enzyme mix and 2X reaction mix.

SuperScript III RT is not significantly inhibited by ribosomal and transfer RNA and can be used to synthesize cDNA from total RNA. Platinum Taq DNA Polymerase High Fidelity is a an enzyme mixture composed of recombinant Taq DNA polymerase, Pyrococcus species GB–D polymerase, and Platinum Taq antibodies, which block polymerase activity at ambient temperatures enabling hot start PCR.

The 2X reaction mix consists of a proprietary buffer system optimized for reverse transcription and PCR amplification, Mg2+ optimized for universal use, deoxyribonucleotide triphosphates, and stabilizers.

The SuperScript III One-Step RT-PCR System with Platinum Taq High Fidelity offers:
• High-fidelity end-point detection and analysis of RNA molecules by RT-PCR
• PCR amplification and cDNA synthesis in a single tube
• Detection of a wide range of RNA targets from 300 bp to 10 kb
• Increased specificity with wide cDNA synthesis temperature range of 45−60°C

Note: For superior one-step RT-PCR performance, we recommend the SuperScript IV One-Step RT-PCR System or SuperScript IV One-Step RT-PCR System with ezDNase. The SuperScript IV One-Step RT-PCR System combines the high processivity of SuperScript IV Reverse Transcriptase with the high-fidelity of Platinum SuperFi DNA Polymerase to provide unmatched product yields, specificity, and sensitivity in less time and for a broad target range, even with suboptimally pure RNA samples.

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 High Fidelity
Quantity100 rxns
Reaction FormatPremixed Components
Reagent TypeReverse Transcription
Reverse TranscriptaseSuperScript III
Ribonuclease H ActivityReduced
Shipping ConditionDry Ice
Size (Final Product)Up to 10 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 High Fidelity Enzyme Mix (100 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.

View all SuperScript™ III One-Step RT-PCR System with Platinum™ Taq High Fidelity DNA Polymerase, 100 rxns FAQs

Citations & References (2)

Citations & References
Abstract
Transcription precedes loss of Xist coating and depletion of H3K27me3 during X-chromosome reprogramming in the mouse inner cell mass.
Authors:Williams LH, Kalantry S, Starmer J, Magnuson T,
Journal:Development
PubMed ID:21471155
'Repression of Xist RNA expression is considered a prerequisite to reversal of X-chromosome inactivation (XCI) in the mouse inner cell mass (ICM), and reactivation of X-linked genes is thought to follow loss of Xist RNA coating and heterochromatic markers of inactivation, such as methylation of histone H3. We analyzed X-chromosome ... More
Influenza A virus molecular virology techniques.
Authors:Zhou B, Wentworth DE,
Journal:Methods Mol Biol
PubMed ID:22528160
Molecular biological techniques for genomic analysis and for creation of recombinant viruses are critical tools in our efforts to understand and combat influenza A viruses. These molecular virology approaches are used in diagnostics, basic research, molecular epidemiology, bioinformatics, and vaccine development. The majority of the techniques used to study this ... More