|Subject: Offer for bulked segregant mapping of C. briggsae mutants|
|From: "Ray Miller" |
|Date: Thu, 2 Aug 2007 12:05:05 -0500|
|To: "'Scott Baird'" |
As announced at the Evolution Workshop at the 16th Int. C. elegans meeting, we are offering to do analysis of bulked segregants for any single-gene mutants you have in C. briggsae. The objective is to increase the density of mutants mapped to the new whole-chromosome assembly CB3. We would like to receive the samples by Friday, August 17. Please pass on this offer to anyone you think might be interested.
The mutants, on the AF16 background, should be crossed to HK104. F2 animals should be phenotyped and sorted into mutants and non-mutants.
We have had good success with this protocol provided by Eric Haag. Pick about 100 mutants into 100 ul of lysis buffer and 100 non-mutants into a second tube of lysis buffer. The lysis buffer includes 50 mM KCl, 10 mM Tris, pH 8.2, 2.5 mM MgCl2, 0.45% NP-40, 0.45% Tween-20, 0.01% gelatin (and 1 mg/ml inactivated proteinase K). Freeze the samples and ship on dry ice to me at the address below.
Raymond D. Miller, Ph.D.
Washington University School of Medicine
Department of Genetics, Box 8232
4566 Scott Avenue
St. Louis MO 63110