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From 2014.igem.org

BYU 2014 Notebook Edit September October

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Week of September 5

Sept 3

(TR,CB) Performed SOEing PCR on the front and back halves of insert to go into pSR47s plasmid. We used the high fidelity q5 DNA polymerase.

Sept 4

(TR) Ran our PCR product from yesterday on a gel electrophoresis to determine if the SOEing worked. The gel came out with only a band at the 500bp range, therefore we know it didn't work. This shouldn't be a problem since we are still waiting to have some N. multiformis grow to be used in conjugation.

Week of September 12

Sept 8

(TR,CB) Ran SOEing PCR again with q5 as the polymerase.

Sept 10

(TR,CB) Ran a gel electrophoresis on Monday's PCR product. Still only a band at the 500bp mark. Reviewed literature on PCR techniques and tried to figure out why our SOEing isn't working. Didn't find anything new.

Sept 11

(TR) Checked on the growth of both the N. multiformis and N. eutropha. Neither one appears to have any growth. We cannot begin our work on N eutropha until be can grow enough to get a template from for PCR to make the front and back ends of our insert to knock out serB from the genome.