cDNA sythesis using Vilo from Invitrogen
Overview
Overview
This protocol describes how to make cDNA from RNA using Invitrogen's ViLo kit. This kit will make single stranded cDNA using first-strand synthesis from single stranded RNA, normally purified using Trizol. The Vilo kit uses random primers (not oligo DT).
Protocol
Protocol
- Add ingredients to PCR tube.
- DO NOT FORGET TO MAKE A NO RT CONTROL. Omit the SuperScript Enzyme mix and replace with H2O
- Gently mix ingredients by pipetting up and down.
- Place PCR tubes in a thermal cycler and run the following program
- 25ºC 10 min
- 42ºC 60 min to 120 min (Longer reaction time will give you more cDNA)
- 85ºC 5 min (to terminate reaction)
- If you started with more than 2 µg of cDNA, the cDNA can be diluted 1:20, which will result in a concentration of ~100 ng/µl.
- Store at -20ºC
Reaction mix