From owner-celegans@net.bio.net Fri Nov 04 22:00:00 1994 Path: biosci!rutgers!sgigate.sgi.com!enews.sgi.com!decwrl!tribune.usask.ca!quartz.ucs.ualberta.ca!elegans.biology.ualberta.ca!user From: Morris_maduro@biology.ualberta.ca (Morris Maduro) Newsgroups: bionet.celegans Subject: injection problems solved Followup-To: bionet.celegans Date: Sun, 06 Nov 1994 12:49:43 +0200 Organization: Genetics U of Alberta Lines: 21 Message-ID: NNTP-Posting-Host: elegans.biology.ualberta.ca Hi fellow worm breeders, I had previously posted a cry for help regarding "injection problems." I received about a dozen replies by e-mail. Aside from my rol-6 recovery being lower than optimal in controls (by about tenfold), the overwhelming consensus on my injection problems was that the DNA had to be clean. Some reported they had never obtained transformants from RNase-treated DNA (even after phenol), and some claimed multiple extractions was the answer. I repeated my mini-prep, avoiding RNase and doubling the organic extractions (2xphenol, 2xphe:chlor, 2xchlor). After a set of injections on Hallowe'en I got 40 F1 rollers. Many thanks to those who sent me suggestions! Morris M. -- Morris Maduro | 'Worms Rule' Dept of Biological Sciences | 'OK' University of Alberta | Edmonton, AB Canada | Morris_maduro@biology.ualberta.ca .