Genomic DNA extraction from mosquitoes
Freeze mosquitoes and separate 1 mosquito per tube .
Add 25 uL of fly grinding buffer .
Grind mosquitoes with a pestle .
Rinse pestle with 25 uL of buffer .
Incubate at 65o C for 30 min .
Add 7uL 8M KAc .
Incubate at ice for 30 min .
Centrifuge at max speed for 15 min at RT .
Collect supernatant and add 0.7 vol ( 28 uL over 40 uL ) of room temperature of isopropanol ( or 1 vol of ice cold 100 % EtOH ) and incubate for 5 min at room temperature .
Centrifuge at RT for 15 min at max speed .
Discard supernatant and add 100 uL 70 % EtOH .
Centrifuge for 5min at max speed at RT .
Discard supernatant and add 100 uL 100 % EtOH .
Centrifuge for 5 min at max speed at RT .
Discard supernatant and aird dry at RT for 1 hour .
Resuspend in TE , heat for 10 minutes and quantify with nanodrop
