DsRNA Preparation protocol (by Ehab Lab)
Prepare the following mix (don’t vortex but mix well)
| Reagent | Amount |
|---|---|
| 10X Transcription Buffer | 10uL |
| RNAse Out | 1uL |
| 10mM NTPs | ~1ug |
| Template DNA | 1uL |
| RNA Polymerase | 4uL |
| H2O | up to 100uL |
- Incubate overnight at 37°C
- Add 2uL DNAse and incubate 10 minutes at 37°C
- Check 1uL on a gel (add Bleach to avoid RNA degradation)
- Purify the dsRNA with phenol/chloroform/isoamylalcohol method.