Does anyone have experience or detailed protocol for dsRNA extraction from HT115 bacteria? I have been using Qiazol to extract total RNA from bacteria, but after a anealing step I do not seem to get any specific bands on a non-denaturing agarose gel. Someone any tips or protocols that you want to share?
Many thanks!
I don’t offhand remember seeing this done? When people want dsRNA for injection or soaking they usually make it in vitro using RNA polymerase, I don’t recall people purifying it from HT115 bacteria.
Indeed, it seems to be the most done for injection/soaking. Why I’m trying this is because dsRNA extractions are easy to upscale/cheaper to perform than in vitro (which easily cost 25-30 euro per reaction).
Thanks for the reply
It does seem though that there are papers out there. The first one I glanced at doesn’t look great; for another one I only read the abstract. Neither one says they do the obvious, high quality test (making sure they get good gene-specific effects and don’t get weird or undesired effects), but if you just want to get dsRNA from HT115 it looks pretty easy, and might even work. There could be worm papers or more careful non-worm papers out there that I didn’t see, I only looked very quickly.