Heat Shock Assay for Positive Control

Model Organism: Worms Methods & Reagents
1

Hi All,

Can someone share their protocol for a heat shock assay? I am looking to do a simple experiment (to serve as a basis for a positive control in a future experiment), where I heat shock the hsp-16.2::GFP strain and quantify the amount of fluorescence. I have anecdotally heard and seen different things in the literature, so would appreciate a clarification. Is the best temperature to carry out heat shock 35C?
Should I pre-warm a free-of-worms, seeded NGM plate at 35C for 30 mins, then transfer 15-20 worms, put the plate with worms back at 35C, wait (30 or 60 mins?) and then measure fluorescence? The critical steps seem to be not to heat shock above 35C and to pre-heat the plate ahead of time rather than placing the plate with worms from RT or 20C directly to 35C because this would allow the worms to acclimate to the increasing temperature on the plate and would not be a true heat shock. Just wanted to double check that that’s right?

Thank you,
B

2

So I went ahead with the experiment today and thought I’d share the results in case anyone else plans to do this. I pre-warmed a seeded NGM plate at 35C for roughly 30 mins, quickly transferred 20 worms from 20C to this plate, heat shocked at 35C for 40 mins and imaged immediately. I observed a robust upregulation of GFP in the hsp-16.2::GFP strain compared to the non-heat shocked control of the same strain.

3

Great, glad it worked! Thank you for sharing your protocol and your success with the forum!