Protocol for Standard RNA Synthesis (NEB #M0378)
| Components |
Amount |
Concentration |
|---|---|---|
| H2O |
X µl |
|
| 10X Reaction Buffer | 2 µl |
|
| NTP | X µl |
0.5 mM each |
| Template DNA | X µl |
0.2–1 µg |
| RNase Inhibitor (optional) |
0.5 µl | 1 U/µl final |
| Fresh DTT (optional) | X µl |
5 mM final |
| T3 RNA Pol | 2 µl |
|
| Total Volume |
20 µl |
Incubate at 37°C for 1 hour. For shorter (< 300 nt) transcripts incubate at 37°C for 2–16 hours.