Hi, I’m going to be running a lifespan assay on worms exposed to RNAi from the Ahringer feeding library.
Normally, I would use FUDR and keep the worms on their same plates. However, the effectiveness of the silencing would obviously decline significantly over the lifespan. The other option is to transfer worms to fresh IPTG-induced lawns every other day or so, which adds the risk of harming the worms. I was wondering if anyone had performed lifespan assays using RNAi and if people had suggestions about the best method to avoid the experimental error described.
Thank you
could you do promoter driven RNAi using a ubiquitous promoter?
I put worms on RNAi plates with FUDR until they have finished laying progeny. They I transfer them to RNAi plates without FUDR. I transfer to fresh RNAi plates about every week, or sooner if the plates become contaminated.
Once they are getting quite old and sick, I leave them on the plates because transferring them can damage them.
I find that the RNAi effect remains strong following this procedure.
i’ve performed RNAi to induce sterility in worms and it looks fine.