Edited by Chang Zhu
1. Grow cells in 96 deep-well block.
2. Pellet cells by centrifugation at 2,000 rpm for 5 minutes.
3. Pour off the supernatant. Drain off the rest of supernatant by inverting the block on
paper towels or by aspiration.
4. Cover the plate and freeze at -80 °C for 15 minutes.
5. Thaw the plate at room temperature.
6. Resuspend the cell pellet in 100 ml P1 + 0.1 mg/ml RNase A.
Vortex at maximum speed for 2 minutes.
7. Add 100 ml P2, vortex at low speed for 10 seconds. Let stand on
ice for 5 minutes. Alternatively, add 100 ml P2, shake by hands to mix, and then
leave at room temperature for 1 hour.
8. Add 100 ml P3, vortex to mix. On ice for 5 minutes.
9. Centrifuge at 3,000 rpm for 30 minutes.
10. Remove 200 ml supernatant and add to
a second deep well plate containing 200 ml isopropanol
in each well.
Precipitate in the deep well plate by centrifuging at 3,000 rpm for 30 minutes.
Alternatively, transfer 200 ml to a microtiter
plate, precipitate DNA. Discard the supernatant and transfer the remaining
200 ml to the microtiter plate with pellets,
11. Wash with 70% ethanol. Decant the supernatant, drain inverted on a
paper towel. Dry under vacuum or air dry.
12. Resuspend in 100 ml ddH2O or TE.
See also Alkaline lysis plasmid miniprep.