microinjection

Beth DeStasio Elizabeth.A.DeStasio at lawrence.edu
Wed Nov 26 21:44:46 EST 1997


Calling successful micro-injectors:

     My lab has hit a brick wall with micro-injection.  We had no problems
getting transgenic lines earlier, but lately we have had no luck.  We are
using
rol-6 as a marker, injecting about 100 ng/ul.  If we use the Shakes &
Epstein
book as a guide we get about 50% sterile animals and no rollers.  When we
decrease the volume of injections, sterility decreases, but still there
are no
rollers in the F1.  We presume the problem is the rol-6 prep.  We've tried
Qiagen preps, alkali lysis, Ctab precipitation.  Does anyone have any
suggestions?  Incantations?  Has anyone tried the endotoxin part of the
Qiagen
kit?  Could it be something in our water?

     Responses can come directly to me at
elizabeth.a.destasio at lawrence.edu.
I'll be happy to post responses.
 
Beth De Stasio
Department of Biology
Lawrence University
Appleton, WI 54911






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