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:
- adding a layer of glass beads (amount of the bottom of a 2 ml tube)
- shake the bacteria on intensity 5 for 5 seconds
- spin down the cells for 2 minutes on 14k rpm on a tabletop centrifuge
- take 500 ul of the supernatant
- 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.