TUDelft/16 October 2008

From 2008.igem.org

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October 16th

New way for cell lysis: FastPrep

We've tried a new method of lysis today with our harvested cells. The FastPrep. A machine which shakes glass beads (d= 1 mm)so vigorously it destroys the cells. We couldn't find a proper E. coli protocol, therefore we used a S. cerevisiae protocol (as the machine is in the yeast group). The machine used was a Thermo Savant FastPrep FP120 Homogenizer. The protocol used was:

  1. adding a layer of glass beads (amount of the bottom of a 2 ml tube)
  2. shake the bacteria on intensity 5 for 5 seconds
  3. spin down the cells for 2 minutes on 14k rpm on a tabletop centrifuge
  4. take 500 ul of the supernatant
  5. perform luciferase and protein content measurements on the supernatant.

Growing cultures for miniprep

We've grown the cultures of our thermoparts in biobrick vectors o/n at 37ÂșC.