What is the optimal reaction temperature for EnzScript M-MLV Reverse Transcriptase RNase H-?

For routine first-stand synthesis, incubation at 42°C is recommended. For RNA templates with difficult regions containing secondary structures or GC-rich regions, incubation temperatures up to 50°C may be used to increase accessibility of EnzScript M-MLV Reverse Transcriptase RNase H– to these regions.

Note: A low temperature incubation step of 25°C for 2 minutes (poly oligo(dT)) or 25°C for 10 minutes (random hexamers) is recommended for primer extension (to increase the primer Tm) before elevating the temperature for first-strand synthesis. 


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