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