Fire hsp-16 based vectors - should strains carrying these as Ex arrays be heat-shocked gradually or quickly, ie. adding to pre-warmed or RT plates
prior to heat-shock? And what temp is/are recommended?
Hi Coltom,
In my experience, each array/gene being heatshocked is different. Some are great with a relatively short, acute shock (37ºC-30min), and some only show a reasonable response with a longer, gentler heatshock (32ºCfor1hr, recovery@25ºC for 30min., re-heatshock at 32). Some heatshock constructs that I’ve had injected are really toxic to the worms (as judged by both few array-bearing animals or really sick ones), and some seem fine.
Sorry if that’s not too much of a help! I know hearing basically “try a bunch of things, who knows what will work for you” isn’t likely what you wanted to hear. If you already have the arrays, and have a relatively easy readout for “success” of your gene’s expression, I would recommend trying a bunch of different methods out. You could look through a few papers that use heatshock and follow a few different published methods (it seems everyone does hs different anyways).
Good luck!
-dylan
Cheers - that’s what I thought.