What do you feel is the most efficient mating technique? I usually pick 10 males and 2 L4 hermaphrodites to a plate with a tiny spot of food and separate them the next day. I get about 75% chance of a successful mating using this method. How can I bump up my success rate?
Thanks,
Mark
I always use 4 to 5 L4 hemaphrodites, mate them at 15oC for 2 days, then single those hemaphrodites to different plates. And only single F1 from those successfully mated plates.
How many males do you use and what if your efficiency?
usually 6-10 males, depends on how many males I can easily get.
Given I single P0, as long as one or two hemaphrodites get mated, that will give me 40-50% males on that plate.
pairwise (1-male-to-1-hermaphrodite) matings of N2 worms on a small spot of OP50 (15 microL grown overnight) typically gives me a mating efficiency of 75%.
depending on your needs and the health of the strains though sometimes it’s better to overload with either males or hermaphrodites.
does anyone get 100% mating efficiency? I think 75% needn’t be improved upon. but if you must, i suggest using the fog-2 background that feminizes hermaphrodites. mating into paralyzed worms will also increase your success rate.
I have found it is most effective using a plate with no food. When you pick the hermaphrodites make sure to grab a little food from their plate with the worms. This most often means the hermaphrodites will not move from the small dot of food that came with them to the plate and the males will look for the hermaphrodites.
I then single the hermaphrodites after one day of mating to see progeny.
I use 2-3x the males as hermaphrodites. To many males can be a problem because they try mating with each other instead of the hermaphrodites.
Guang, I tried your method of mating 2 days at 15C and noticed a huge increase in mating efficiency (to over 75%). Thanks!
-Mark