M-MLV Reverse Transcriptase Buffer

While the volume is dependent on the starting amount of RNA used for the first-strand synthesis and the abundance of the target gene, we'd recommend starting with 10% of the first-strand reaction for your PCR reaction.

The optimal temperature for for MMLV is 42 degrees C. The optimal temperature for SuperScript II RT is 42 degrees C, and can be used up to 50 degrees C. The optimal temperature for SuperScript III RT is 50 degrees C, and can be used up to 55 degrees C. For some qRT-PCR reactions where gene-specific primers are used, you can do the RT reaction at 60 degrees C. The optimal temperature for SuperScript IV RT is 50 degrees C, but can be used up to 65 degrees C.

Moloney Murine Leukemia Virus (M-MLV) Reverse Transcriptase is not well-suited for elevated temperatures. Typically, M-MLV RT operates optimally at temperatures around 37 degrees C to 42 degrees C.

To perform reverse transcription at elevated temperatures (e.g., 50 degrees C or higher), consider using thermostable reverse transcriptases such as SuperScript IV or Maxima H Minus, which are engineered to perform efficiently at higher temperatures.