Team:UCLA/Notebook/Honeybee Silk/19 May 2015

5/19

Cell Harvesting

  • Cells were grown up in 250 ml of LB medium and induced with IPTG. See 5/18 for details.
  • The Cells were split up into 5 pre weighed 50 ml falcon tubes, with 45 ml of cells in each tube.
    • I should have taken the OD 600 at this point, but I forgot to.
  • Mass Balanced the tubes and added a water tube, and spun down the cells at 5300 rpm at 4 C for 15 min. to pellet the cells.
  • Decanted the supernatant.
  • Weighed the pellets
    • In total, the wet weight of all the pellets across all 5 falcon tubes was 2.42 grams, approximately 0.5 grams per 45 ml of culture.
  • Froze down the pellets in the falcon tubes at -80C
  • Next up, will lyse the cells and collect protein with BugBuster


Transformation