IGEM:VGEM/2007/Notebook/2007-6-14

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June 14, 2007

  • Amy- paper presentation
  • Discussion of wireless repressilator
  • Confirmation of final melanin DNA sequence
  • Practice miniprep
    • 1. Add 1.5 mL each of transformed cells in LB broth to three centrifuge tubes
    • 2. Spin the cell-broth mixtures down
    • 3. Resuspend cell pellet in 250 ul of buffer P1 by vortexing
    • 4. Add 250 ul of buffer P2. Invert until solution becomes viscous and clear
    • 5. Add 350 ul of buffer N3. Invert 6 times.
    • 6. Centrifuge for 10 min at 13,000 rpm
    • 7. Pipette out supernatant
    • 8. Centrifuge for 1 min and discard flow-through
    • 9. Add 0.5 mL of PB buffer
    • 10. Centrifuge for 1 min and discard flow-through
    • 11. Repeat steps 9 and 10
    • 12. Add 0.75 mL of buffer PE and centrifuge for 1 min
    • 13. Discard flow-through and centrifuge for 1 min
    • 14. Add 50 mL of elution buffer and wait for 1 min
    • 15. Centrifuge for 1 minute and place in freezer
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