I'm having trouble getting males from the C. elegans strain hsp-4::GFP (SJ4005). I tried HS submitting L4 animals to different temperatures and times and also him-14 RNAi and neither approach worked (I did the same with N2 to see if would work and it worked, both approaches, so my problem is with this specific strain). Has anyone had this problem with this strain before? Does anyone have any suggestions of what I can try?
I did heat shock for this strain and had no issues. I found with the HS you need robust L4's with visible vulva patches and for the heat shock, I would leave them in for 2-3 days at 25C and would have plenty of males to start a breeder plate. Hope this helps.
Skylar King so you heat shock them at 25°C? How many L4 do you usually incubate on each plate? I've never tried temperatures below 30°C for HS, I'll try this and see if it works. Thank you for your help!
I find heat shocking whole plates with a large number of L4s at 30°C for 6h a day for 2-3 days always gets some males.
I generally just put 5-8 robust L4's on a 6cm dish. You could try several plates. I had some temperature sensitive animals in the past so found that 25C worked well for those. Hope this helps!
Skylar King yes!! I did just the way you said and it worked! I got enough males to start a breeder plate. Thank you so much for your help! :)
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