Miniprep Protocol 12/21/00

Grow one colony in 3ml LB + antibiotic overnight (>6hr) at 37C shaking

1.  transfer 1.5ml to eppendorf

2.  Spin 8000rpm x 3 min

3.  discard supernatant

4.  resuspend pellet in 100ul of Resuspension Solution

5.  (incubate 5 min at rt)

6.  add 200ul cell lysis solution, mix by inverting

7.  (incubate 5 min on ice)

8.  add 150ul Neutralization Solution and vortex

9.  (incubate 5 min on ice)

10. spin max rpm x 3 min

11. take 350ul of supernatant and transfer to fresh eppendorfs

12. add 700ul EtOH

13. incubate 2 min at RT

14. spin max rpm x 3 min

15. spill out supernatant and gently apply 70% EtOH wash.

16. spill out supernatant gently.

17. dry eppendorfs by suspending upsidedown on a paper towel in incubator

18. resuspend pellet in 30-40ul ddH2O and keep on ice.