I know that daf-2 (e1370) is temperature sensitive and should be maintained at 20. Does anyone know if that precludes me from heat-shocking for males? Our protocol is 31 degrees for 5hrs on L4s. I’m wondering the same about the double daf-16 (mu86); daf-2 (e1370).
Thanks,
Kris
Usually, we maintain a mating plate of daf-2 males but it can’t hurt to try it. Your first progeny might be dauers. If you are really worried about it you could maintain the animals on daf-16(RNAi) prior to heat shock. Double mutants should be fine to heat shock. Just keep in mind that animals without daf-16 are are compromised for thermotolerance (in our hands we get ~50% mortality of young adults after 6-8 hours at 35C, so you should alright with 5 hours at 31C). Good luck.
-A.