Quick genomic DNA prep for single flies
used in Orgogozo et al., 2006
(based on Gloor et al., 1997)

very fast, good to amplify fragments smaller than 1kb; this genomic DNA can only be kept for a few months at –20C. Use another protocol if you want to amplify larger fragments or keep the DNA longer.

squishing buffer (100mL):       1mL Tris Hcl 1M pH=8.2
                                                200uL EDTA 0.5M
                                                500uL NaCl 5M
                                                94mL milliQ water
    autoclave. (1mL proteinase K and 2.5mL RNAse to be added)

1. put the fly in an Eppendorf tube and then freeze at -80C. Flies can also be stored in acetone at RT for at least a year; in this case, remove acetone and wait about 3min (not more than 3min, to prevent tissues from drying and solidifying).
2. prepare the solution: 100uL squishing buffer+1uL proteinaseK (20mg/mL) +5uL RNAse (20mg/mL). 50ul is needed for each fly.
3. grind the fly in 50uL with disposable tissue grinder.
4. incubate the squished flies at 37∞C for 25min.
5. inactivate proteinase K at 95∞C for 1-2min. Then put on ice.
6. centrifuge 1min at 13.000rpm.
7. put the supernatant (52uL) into a new tube. Store at -20∞C.

use 1uL for each 25uL-PCR reaction.