Ultra-fast alkaline lysis plasmid extraction
Robert S. Cormack and Imre E. Somssich, Technical Tips Online
- 0.2 ml o/n culture in eppie
- add 0.2 ml solution II (1% SDS, 0.2 N NaOH), invert 4 times
- immediately add 0.2 ml solution III (3 M potassium acetate, pH 5.5), mix by inversion
- centrifuge at 14 krpm for 2 min
- transfer supernatant in eppie with 0.5 ml 100 % isopropanol, mix by inversion
- centrifuge for 1 min, discard supernatant
- spin again briefly to collect residual isopropanol, remove with a pipette
- dry briefly
for screening, resuspend pellets directly in 10 ml restriction digest mixture, load all on gel