TUDelft/16 October 2008

=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 37ºC.