I’ve been trying to use RNAi against him-14 or him-3 to make males, however the efficiency is extremely low especially in my mutant background. Does anyone have any tips or tricks to increase efficiency? Can I play with IPTG concentration or amount/concentration of bacteria lawn?
We’ve had success with the him-14 RNAi plasmid from the Hubbard lab. Most of the males are produced on the last day of egg-laying. We initiate RNAi feeding as L1 larvae; when they reached adulthood, we transfer individual hermaphrodites each day to fresh RNAi feeding plates. We’ve seen 10-15% males on the last plate of progeny.