tail DNA extraction

Prepare Genotyping Lysis Buffer:

50mM TRIS-HCl ph8, 10mM EDTA, 100mM NaCl, 1%SDS

- digest o/n at 55°C in 665µL Lysis Buffer with 35µL Proteinase K 10mg/mL (f.c. -> 500µg/mL)

- Add 700µL P:C:I, vortex, spin max speed benchtop centrifuge for 10' 4°C

- Transfer 600µL of the acqueous phase in new eppendorf

- Add 600µL P:C:I, vortex, spin max speed benchtop centrifuge for 10' 4°C

- transfer 500µL of acqueous phase in new eppendorf

- add 1mL Et-OH 100%

- incubate 20' RT

- spin max speed 30' at 4°C

- discard supernatant and add 300µL Et-OH 70%

- resuspend in H20 (50-100µL depending on the pellet)