today i tried in vitro fertilization for my fish for the 3rd time. still didn't work. how hard is it to fertilize a bunch of eggs with the sperms right?

but i saw the most amazing thing under the microscope at 80x mag. the sperms are inactivated at high salt concentration (hank's balanced salt solution with 4.2 mM sodium bicarbonate) so they just looked like tiny dots not moving. once i added egg water into the solution, they instantly got activated and started swimming. cutest thing in the world! i could hardly see their tails, but i would think they have them...

which means the sperms i collected were fine. that means the eggs were bad. fine. i'll get up early tomorrow, i mean today, morning to do the dissection at the egg's prime time. well... sighz..why is there such a thing as biorhythm!!

and i don't suppose any of you guys sequence fosmids of 40 kb size right? just in case you do.. how do you yield results with fosmid sequencing? it just doesnt' work. i've tried different primers. maybe i need to digest it. anybody?