Miniprep (McGill)
From 2006.igem.org
1. Pour 1.5 mL of overnight seeding broth into 1.5 mL Epi tube.
2. Spin in centrifuge at max speed for 1 min. Decant supernatant.
3. (Pour in new broth and spin again to increase pellet size if necessary).
4. Add 250 uL of Buffer P1 (refridgerated) to each tube. Vortex until cell pellet is completely resuspended.
5. Add 250 uL of Buffer P2 to each tube, inverting gently 4 to 6 times after addition. Allow tubes to sit for 4 to a maximum of 5 minutes.
6. Add 350 uL of Buffer N3 to each tube, inverting gently 4 to 6 times immediately after addition.
7. Centrifuge tubes for 10 minutes at max speed.
8. Transfer supernatant to QIA prep spin column by pipetting or pouring, being careful to avoid transferring the cell debris which can clog the column.
9. Centrifuge spin column for 1 min, discard flow-through.
10. Add 750 uL of Buffer PE to each spin column.
11. Centrifuge for 1 min and discard flow-through.
12. Centrifuge for another minute, again discarding flow-through (this removes any residual PE buffer).
13. Transfer spin column to 1.5 mL Epi tube.
14. Add 50 uL of dH2O to center of column filter. Allow columns to sit for 1 min.
15. Centrifuge columns for 1 min. Preserve the eluate, which is the miniprepped DNA sample.
NB. To wash spin columns, fill with 500 uL of dH2O, spin for 1 min and discard flow-through. Repeat with another 500 uL of dH2O, spinning for another minute. Columns may be washed like this and reused up to 5 times.
