IGEM:Peking/2007/count:Miniprep

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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
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