Editing Purification of VLP (section)

== Culture and Harvest Cells ==

1, Streak a VLP clone in Nico21(DE3)pLysN&nbsp; onto a LB/KC plate, label the plate (write clone #), grow at 37 oC incubator O/N, agar side up

2, Wash E coli into an autoclaved glass culture&nbsp;tube with&nbsp;15 ml LB/KC medium

3. Culture cells for 1 hour at 37 oC in a shaker incubator

4. Pour 15 ml the above&nbsp;starter stock into 1 L LB/KC medium, add 20 ml of 50X 5052 using autoclaved 100 ml glass graduated cylinder

5. Grow cells to OD600&nbsp; 0.6 ~ 1.0

6. Add 0.5 ml 1 M IPTG, induce at 37 oC for 4 to 6 hours. You can also leave the&nbsp;induction overnight '''providing you culture&nbsp; the cells overnight at 16 oC in the cold room downstairs'''. Sometimes cells may be lysed during overnight culture. So it would be better to spin down cells after 4 hours induction.

7. Spin down cells, 5 min at 5000 rpm using a high speed centrifuge.

8. Combine cells into one centrifuge tube, use LB/KC medium (supernatant liquid from previous spin) to wash cells into one tube. Spin again to collect pellet.

9. Record wet cell weight. Label tube

10. Store cells (tube) at -80 oC freezer for 1 hour&nbsp;or overnight.