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 this kit:
• Sensitive—routine detection down to 0.01 pg total RNA (see figure)
• Convenience—one-step format for speed, convenience, and less reaction-to-reaction variability
• Specificity—uses SuperScript® III RT for cDNA synthesis up to 55°C, for more specific priming with gene specific primers (see figure)
• 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.
Contents & storage
• 50 µL SuperScript® III RT/Platinum® Taq Mix
• 1 mL 2X Reaction Mix (containing 0.4 mM of each dNTP, 3.2 mM MgSO4)
• 500 µL 5 mM Magnesium SulfateStore all components at -30°C to -10°C.