User:Robert J. Schiemann/Notebook/Lipsick Lab/2010/02/02

Drosophila melanogaster Genetics

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Spermatogenesis Dissect gfp-PCNA x tdt-Myb Flies Chose orange eyed males Dissect testes in 1x PBS Solution. Store in eppendorf with 1x PBS until dissections are finished. Fix with 4% Formaldehyde PBS Solution 30 minutes Wash 3 x 5 minutes in 1x PBS-T Incubate 30 minutes with PBS 0.6% Triton. Wash once with 1x PBS-T Stain with ToPro3 1:1000 in 2% NGS 30 mins at RT Wash 4 x 5 minutes in PBST Mount in Vectashield Collect Flp-out virgins in case PCR fails again Re-run PCR with new reagents