2014hs.igem.org/team:Charlottesville RS/Notebook/041414

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April 14th, 2014

Alli Ambrosini

The results of the transformation efficiency kit and John’s ligation procedure were ready today, and they were very different. There was no growth, but the ligation results were very good. The cells were competent. There were are several possibilities as to why our part did not work in the transformation efficiency kit, including:

- There was no DNA to begin with due to a mistake in procedure

- Lauren and Alli incorrectly measured the volumes while pipetting during the lab- John’s group just might have gotten lucky and got all the competent cells, since they were all taken from the same tube

- The DNA was not thawed enough, and might not have picked up any DNA.

Results:

-DNA/Chlor. : transformation control = growth

-Parts A & B from iGEM : ligation control = growth

-Transformation Efficiency Kit : our part = no growth

For This Week:

1. Rehydrate DNA and transform into E.Coli to multiply DNA

2. Miniprep

3. Digest

4. Ligate

5. Transform

Alli Ambrosini, Lauren Ewell

Took bacteria from John’s “NEB10 beta” plate from 4/8/14 and streaked it onto the SOB agar plate in hopes of getting more bacteria.

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