From owner-celegans@net.bio.net Wed Nov 26 22:00:00 1997 Path: biosci!biosci!not-for-mail From: Beth DeStasio Newsgroups: bionet.celegans Subject: microinjection Date: 26 Nov 1997 18:44:46 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 27 Sender: daemon@net.bio.net Approved: wmorgan@acs.wooster.edu Distribution: world Message-ID: <65imqu$qhr@net.bio.net> NNTP-Posting-Host: net.bio.net 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@lawrence.edu. I'll be happy to post responses. Beth De Stasio Department of Biology Lawrence University Appleton, WI 54911 .