< Miniprep(Difference between revisions)
|back to protocols|
TENS method of minipreps. Fast but dirty.
- Transfer 1.5 mL of an overnight culture containing your plasmid to an eppendorf tube and spin at 5000 rpm for 5 min in a tabletop centrifuge to pellet the cells.
- Remove and discard the supernatent.
- Resuspend either in 50 μL of LB or P1 buffer or TE/RNAse
- Add 300 μL TENS buffer
- Mix by inverting 5 times.
- Add 100 μL 3M NaAc pH 5.2
- Mix again
- Spin down at top speed for 5 minutes
- Transfer supernatant to a new tube (pouring works)
- Add 1 mL 100% EtOH (ideally ice cold)
- Spin down at top speed for 5 minutes (ideally at 4˚C.)
- Wash with 0.5 mL of 70% EtOH
- Pour out the supernatant
- Spin again to find the rest of the supernatant
- Air dry for 10 minutes
- Resuspend the dried pellet in 30 μL of water or TE buffer.
- Do your molecular bio
- 10 mM Tris-HCl, pH 8
- 1 mM EDTA
(500 mL recipe)
- 5 mL 1M Tris-HCl pH 8.0
- 12.5 mL 20% SDS
- 5 mL 10N NaOH
- Enough H20 to 500 mL