User:Robert J. Schiemann/Notebook/Lipsick Lab/2009/04/14

Drosophila melanogaster Genetics

Main project page Previous entry      Next entry

Other Met with Heather about plans for week. All days Virginize and create line for control cross. Wenesday Dissections of approximately 20 males from [ubi-RFP] and [GFP-myb, RFP-H2Au] lines into PBS. Fix 30' in 4% Formaldehyde, PBS-T at RT. Wash 4x5' Block in 2% NGS O/N Thursday Change NGS. [ubi-RFP] Stain 30' in Totpro 1:500 Wash 3x10' or 4x5' in PBS-T Mount immediately. [GFP-myb, RFP-H2Au] Incubate w/ 1:5 dilution of a-Cyclin B (Mouse Antibody) O/N. Friday [GFP-myb, RFP-H2Au] Wash 3x15' Incubate w/ 1:500 dilution of a-Mouse IgG 633nm (Goat antibody) for 2 hours. Wash 3x15' in PBS-T Mount Following Week Vizualize both Spermatogenesis Collected a few virgin females of [MH107,hsFLP/FM7Ag ; mRFP, myb+, FRT42D/CyoAg] Some had lost CyoAg, but will not affect the cross. FRT/RFP Recombination Focus off of project for a while. Will continue to maintain stocks.