Quick and dirty PL prep from E.coli

  1. Spin down 2-3 ml induced E. coli culture at 8000 rpm for 1 min

  2. Resuspend with 300 µL of Qiagen P1 buffer (with RNase1)

  3. Add 300 µL of Qiagen P2 Buffer, invert 10 times

  4. Incubate at room temp for 2 minutes

  5. Add 300 µL of Qiagen P3 Buffer, invert 10 times

  6. Centrifuge at 12000 rpm for 5 min

  7. Transfer all the supernatant to a new 2 ml tube

  8. Add 900 µL isopropanol, mix by inverting

  9. Spin down at 12000 rpm for 5 min

  10. Discard the supernatant

  11. Wash with 500 µl of 70% EtOH

  12. Spin down at 12,000 rpm for 1 min, discard the supernatant

  13. Air dry

  14. Dissolve the DNA with 30 µL TE buffer

  15. Spin 12000 rpm, 1 min, and transfer all the clear supernatant into a new 1.5 ml tube.

  16. Load 2 µL on agarose gel to determine the DNA size and concentration

  17. DNA is ready for BAC library prep