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

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Drosophila melanogaster Genetics
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


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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.


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