TUDelft/7 October 2008

From 2008.igem.org

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October 7th

Sonication optimization

To optimize the new use of sonication for lysing cells, we'll be trying the effect of different lengths of sonication on our protein content and luciferase activity. We'll be trying a series of:

  • 1 to 5 times 15 seconds sonication, with 15 seconds pause.
  • 1 to 4 times 30 seconds sonication, with 30 seconds pause.
  • 1 times 60 seconds sonication.

Because earlier luciferase measurements weren't done in duplo, this time we've tried this to see whether it's necessary for following experiments.

Results are displayed in figure 1.


TUDelft071008.jpg Figure 1: Luminescence corrected for protein content (figure 2) of the various protocols of sonication


TUDelft071008 2.jpg Figure 2: Protein content of the various protocols of sonication

It can be seen longer sonication releases more protein, but luciferase gets inactivated after a certain amount of sonicating. We'll use the 2x 15 seconds protocol for our following experiments.

Miniprepping

We've miniprepped K115030 and K115033. K115035 didn't grow o/n, so we'll try to grow it tomorrow on Kanamycin instead for prepping Thursday.