What part of the promoter to clone - (Jul/10/2013 )
My advisor is asking me to amplify the promoter of my gene of interest to be clone into a luciferase vector so we can test the expression under specific conditions. The problem is that I'm not sure how much or what to amplify. Does anyone have any suggestions? Thanks in advance!
It depends. If you want to have guaranteed promoter activity which recapitulates endogenous promoter, the longer the better. I would say 1-3 kb should be fine. You should also include some sequence (50 - 100 bp) downstream of the TSS.
Thanks for your help! I will keep what you suggested in mind.