Heat Treatment Method for P{hs-hid} Stocks

P{hs-hid} expresses hid under the control of Hsp70 regulatory sequences. Consequently, any progeny inheriting a P{hs-hid} insertion can be killed by incubation at 37 degrees C. P{hs-hid} insertions on the Y chromosome are useful for eliminating male progeny during development and P{hs-hid} insertions on balancers are useful for eliminating balancer heterozygotes.

Selected P{hs-hid} stocks are listed among the Dominant temperature-sensitive lethal stocks and a full list of stocks with Y-linked P{hs-hid} insertions is provided here.

We have used the Y-linked P{hs-hid}Y insertion extensively for virgin collection and have found the following approach to be quite reliable.

Allow females to lay eggs in vials. After sufficient time to allow eggs to hatch to larvae, place the vials in a 37 degree C water bath for 1.5 hours. (Make sure the water comes at least halfway up the sides of the vials so that larvae can't crawl up to get away from the heat.) Typically, we allow females to lay eggs for 2 to 3 days and then heat shock sometime on days 4 to 6.

We have found one heat treatment to be sufficient, but we have also heat treated vials for one hour on three successive days as an added precaution.

Our experience is that heat treatment in a water bath is absolutely essential for adequate heat transfer. Our attempts to heat shock vials in a 37 degree incubator were miserable failures. We strongly recommend against trying to heat shock in an incubator. (Don't blame us if it doesn't work. We warned you...)

We have had mixed success heat treating bottle cultures. Vial cultures work so well that we avoid bottles.