Re: Help!!! BAC DNA isolation...# Biology - 生物学
s*l
1 楼
1.start from 3-4ml fresh overnight culture, pellet cell in 8000rpm in cold for
3min
2.resuspend in 200ul P1
3.lysis with with 400ul P2 <5min in RT
4.neutralize with 300ul P3 on ice for 5 min
5.spin down 10min at maxi spped in cold
6.supernatant to a new tube
7.add 0.75ml cold isopropanol
8.spin at maxi speed RT for 15min
9.discard supernatant, wash with 0.5ml ethanol, spin 5min maxi speed in cold
10. asirate supernatant and air dry DNA till pellet starts to turn translucent
Caution: DON'T OVER D
3min
2.resuspend in 200ul P1
3.lysis with with 400ul P2 <5min in RT
4.neutralize with 300ul P3 on ice for 5 min
5.spin down 10min at maxi spped in cold
6.supernatant to a new tube
7.add 0.75ml cold isopropanol
8.spin at maxi speed RT for 15min
9.discard supernatant, wash with 0.5ml ethanol, spin 5min maxi speed in cold
10. asirate supernatant and air dry DNA till pellet starts to turn translucent
Caution: DON'T OVER D