Typical cDNA Synthesis Protocol with WarmStart® RTx Reverse Transcriptase (Glycerol-free) (NEB #M0439)
Incubate the following reaction for 5 minutes at 25°C for annealing and 10 minutes at 55°C for synthesis.
|
Components |
Final Concentration |
|---|---|
|
10X Isothermal Amplification Buffer (Lyo-compatible) |
1X |
|
dNTP Mix (10 mM) |
0.5 mM |
|
Random Primer Mix |
6 µM |
|
RNase Inhibitor, Murine |
20 units/reaction |
|
WarmStart RTx Reverse Transcriptase (Glycerol-free) |
3.75 units/reaction |
|
Nuclease-free Water |
to 20 µl |
|
Total Reaction Volume |
20 µl |
General Guidelines:
- Dilute the WarmStart RTx Reverse Transcriptase (Glycerol-free) (NEB #M0439) to 15,000 U/ml in 1X Isothermal Amplification Buffer (Lyo-compatible).
- Heat inactivate WarmStart RTx Reverse Transcriptase by incubation at 80°C for 10 mins.