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

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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.