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

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