18 (May 18, 2011 GDS)
From 2011.igem.org
TENS How-To:
- 4.5 TE
- 250λ 10% SDS
- 250λ 2M NaOH
Mini-Preps (How-To)
1. Spin 1.5 mL of overnight culture for 30 secs. in micro-centrifuge.
2. Aspirate off all but 200uL of supernatant and resuspend the pellet by vortexing.
3. Add 300uL of TENS and mix by inversion. The solution should become viscous.
4. Add 150uL of sodium acetate and vortex. A fine white precipitate should form.
5. Centrifuge for 2.5 minutes.
6. Transfer the supernatant to clean tube and add 2 volumes (1mL) of room temp ETOH.
7. Mix and pellet DNA by centrifugation for 2-5 min.
8. Wash pellet with 70% ethanol and allow pellet to dry.
9. Resuspend the pellet in 30uL TE.
10. Digest 5-10uL as usual.
- Minipreps (2 each) for:
- pNICE SA-15λ TE
- J024430 GE-15λ TE
- Trans002-5, AB-Resp 30λ
- Trans003-W1 SD, SA-