Description

RNA targets from 100 bp to >12kb can be detected with this system. The amount of starting material can vary from 1pg to 5µg of total RNA. 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 is used to synthesize cDNA at a temperature range of 42° to 55°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 may be used to synthesize first-strand cDNA from a total RNA preparation.

• Using the SuperScript III First-Strand System: cDNA synthesis is performed in the first step using either total RNA or poly(A)⁺-selected RNA primed with oligo(dT), random primers, or a gene-specific primer
• In the second step, PCR is performed in a separate tube using primers specific for the gene of interest
• For the PCR reaction, one of the following DNA polymerases is recommended
• Platinum Taq DNA Polymerase provides automatic hot-start conditions for increased specificity up to 4kb
• Platinum Taq DNA Polymerase High Fidelity provides increased yield and high fidelity for targets up to 15kb
• Platinum Pfx DNA Polymerase provides maximum fidelity for targets up to 12kb

• Optimal reaction temperature: 50°C
• Sufficient for 50 reactions, based on a 20µL reaction size

PCR & Real-Time PCR, Reverse Transcription

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