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

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


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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


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