Dsrna Preparation

Apr 18, 2021

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
  1. Incubate overnight at 37°C
  2. Add 2uL DNAse and incubate 10 minutes at 37°C
  3. Check 1uL on a gel (add Bleach to avoid RNA degradation)
  4. Purify the dsRNA with phenol/chloroform/isoamylalcohol method.