Reaction Conditions (E5315)
- First strand cDNA synthesis can be carried out between 42°C to 55°C. As a starting point, we recommend incubation at 48°C for 15 minutes. Short targets can be completed with shorter incubation times. Longer incubations up to 1 hour may increase the cDNA yield.
- The pre-denature step at 94°C for 1 minute is sufficient to inactivate ProtoScript II Reverse Transcriptase.
REACTION COMPONENTS FINAL CONCENTRATION Reaction Mix 1X Enzyme Mix 1X RNA Template 1 μg–100 pg of total RNA
1 pg–100 ng of mRNA
Gene Specific Primers 100–800 nM