zfish promoters

Geoffrey Doerre gdoerre at mail2.sas.upenn.edu
Sat Jan 21 13:11:30 EST 1995


Russell Buonor/Anatomy (BUONOR at JEFLIN.TJU.EDU) wrote:
: Hello Buono Here.   While its true that after electroporation of zfsh 
: embryos very few cells are found expressing the introduced construct, my 
: original data reported only that dot blot positive animals could be 
: obtained.

i owe russ an apology here.  i didn't mean to sound so irreverent in my 
post, and to be accurate, i didn't try to exactly _repeat_ his results.  i 
used a different pulser (BTX), a different promoter (CMV) and a different 
reporter (lac Z.)  but, after numerous attempts at the lowest settings, 
even using only one pulse, and cleaning the embryos very meticulously, 
survival was below 10%.  and nothing ever expressed, whereas we know from 
microinjections that the construct works.

i didn't mean to imply anything beyond the fact that in my inexperienced 
hands, i got nuthin' but a lot of frustration.  but i'm always 
tempted to try this again when someone brings it up, because it just seems 
like it *should* work.

: I'm happy to see some people 
: are still exploring options in electroporation.  I noted at CSH a 
: group who were using more sophisticated equipment to obtain results.  

me too; keep us posted.
geoff | | | | | | | | | | | | | | | | | | | | | | | ||||||||||||||||||||||
|||||| | | | | | | | | | | | | | | | | | | | | | | | gdoerre at sas.upenn.edu




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