Why does the RobusT™ RT-PCR Kit use AMV Reverse Transcriptase rather than MMLV Reverse Transcriptase?
RNA transcripts exhibiting significant secondary structure must be denatured for efficient reverse transcription. In order to minimize secondary structure, the first strand cDNA synthesis must be carried out at an elevated temperature. While the maximum recommended temperature for MMLV Reverse Transcriptase is 42 °C, AMV Reverse Transcriptase is active at 60 °C in the optimized RobusT™ Reaction Buffer. Thus, the use of AMV Reverse Transcriptase eliminates many problems associated with secondary structure.
