IGEM:Peking/2007/count:Miniprep

Mini Plasmid Purification

according to Transgen Kit Potocol.

1. take 1-4mL over night cell culture ,13000rpm centrifuge for 1 min.discard the supernatant .
2. add 250uL solution RB(contain RNaseA),mix violently.
3. add 250uL solution LB，mix gentlely up and down for 4-6 times,此时溶液变粘，清亮,go to step 4 as soon as possible。
4. add 350uL solution NB,mix gengtlely for 5-6 times.
5. can see white precipitation.
6. 13000 rpm centrifuge 5 min
7. absorb supernatant into Spin column
8. standing for 30 secends
9. 13000 rpm centrifuge 1 min
10. discard the flow-through,add 650 WB,
11. 13000rpm for 1 min,discard the flow-through
12. 13000 rpm for 2 min(in order to wipe out WB(contain ethanol),discard the spin column
13. put the collection tube into a clean EP tube.add 30-50uL dH20(60C) in the center of the center of the collection tube.
14. standing for 1 min.
15. 13000rpm for 1 min.
16. store the DNA solution at -20C