Haynes Lab:Notebook/Synthetic Biology and Bioinformatics for Predictable Control of Therapeutic Gene2/2012/08/22

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QiaPrep Spin

  1. Transfer 2mL of culture to round small round bottom tubes
  2. Divide plate and label (this will be your back up culture)
  3. Drag tip from tube lightly across the agar. Put the tip back in the tube. (Use forceps)
  4. Centrifuge for 3 min (make sure to balance and keep centrifuge at top speed)
  5. Dump waste from tubes
  6. Resuspend pellet in 250uL of PI buffer then vortex
  7. Add 250uL of P2 buffer and invert 6x
  8. Add 350uL of N3 buffer and invert 10x
  9. Centrifuge for 10 min
  10. Label column collection tubes (blue) and dump you centrifuged liquid in columns
  11. Spin for 1 min and dump waste
  12. Add 750uL of PE buffer
  13. Centrifuge for 1 min, dump, centrifuge for 1 min
  14. Place liquid in 1.5mL microcentrifuge tube and add 75uL of EB buffer directly above the white filter
  15. Centrifuge for 2 min toss blue column


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