mating into vulvaless animals

Tritech_Research at lamg.com Tritech_Research at lamg.com
Wed Oct 29 10:06:41 EST 1997


Thomas Burglin writes:
Hi everyone.
We have a putative knock-out, but the animals look vulvaless
and bag.
We need to backcross this.

What is the current state-of-the art for mating
into vulvaless animals?

I remember people mention poking holes into
the hermaphrodite to allow male mating.
What's the best way to do this?

Also, how about injection sperm directly?
Is this done these days? What protocol to make sperms?


Thanks everyone in advance.
Thomas
------------
I don't know about state of the art, but I was able to mate a new
lin-29 mutant (100% blip vulva, 0% mating and egg-laying)
after perforating the vulva area by hand under the dissecting scope
using a hand-held microinjection needle.  Do several, and put them
on a plate with lots of N2 males.  I tried artificial insemination, too.
That was worse, because you have to break off the needle to get the
tip big enough to suck up sperm, and then it was too blunt and
did too much damage to the p-vul recipient.

Andy Papp
tritech_research at LAMG.COM
PS: If the intestines explode out, you have overdone it 8 )
----------
PPS to Bill Morgan:  This arrived with daemon at net.bio.net as the From
and nobody at net.bio.net as the CC.  To work most effectively with most
mailers, it would be better if the From was the actual author and the
CC was celegans at net.bio.net.  That makes it easy for people to reply to
only the author, the list, or both.
Keep up the good work,
Andy






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