ThermoScript™ RT-PCR Systems

Successful RT-PCR of Difficult RNA Templates
ThermoScript™ is an avian reverse transcriptase (RT) designed for high performance with difficult RNA templates. The enzyme was derived from cloned avian myeloblastosis virus (AMV) with point mutations that reduce endogenous RNase H activity and provide increased thermal stability up to 70°C (Figure 1), longer cDNA transcripts, and higher cDNA yields. Combined with PCR enzymes, the ThermoScript™ RT-PCR Systems provide optimal first-strand cDNA synthesis followed by PCR in a convenient, reproducible process.

ThermoScript™ Reverse Transcriptase is available in a variety of formats:

• ThermoScript™ Reverse Transcriptase
  Contains ThermoScript™ RT enzyme and reaction buffer
• ThermoScript™ RT-PCR System for First Strand cDNA Synthesis
  Contains ThermoScript™ RT enzyme, reaction buffer, dNTP mix, oligo dT and random primers, RNaseOUT™ ribonuclease inhibitor, RNase H and DEPC-Water
• ThermoScript™ RT-PCR System with Platinum Taq DNA Polymerase
  Contains the ThermoScript™ First Strand cDNA Synthesis kit above plus Platinum Taq DNA polymerase and PCR reaction buffer
• ThermoScript™ RT-PCR System with Platinum Taq DNA Polymerase High Fidelity Contains the ThermoScript™ First Strand cDNA Synthesis kit above plus Platinum Taq DNA polymerase High Fidelity and PCR reaction buffer

Figure 1: High cDNA yields at elevated temperatures



cDNA was synthesized from total HeLa RNA with ThermoScript™ RT and oligo(dT) primers. 35 PCR cycles were performed with Platinum® Taq DNA polymerase and primers for a 532 bp phosphatase PP2A target. Yield was maintained as the temperature was increased.