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

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April 14th, 2014

Alli Ambrosini

The results of our transformation efficiency kit and John’s ligation thing were ready today, and they were very different. There was pretty much nothing on the efficiency kit, but the ligation results were very good. We got competent cells! We think there are a whole lot of reasons that our part didn’t work in the transformation efficiency kit, including possibly:

- We messed up in the beginning and never had any DNA at all - Lauren and Alli messed up the volumes while pipetting during the lab - John’s group just might have gotten lucky and got all the competent cells, since we took them all from the same tube - The DNA wasn’t thawed enough, and we might not have picked up any

Results:

-DNA/Chlor. -Transformation control = growth RFP Antibiotic Resistance

-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 - rip open E.Coli to get purified DNA 3. Digest - cut up the parts we need 4. Ligate - stick em back together 5. Transform (into giant robots)




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.