Week of 8/26-8/30

Having ran linear PCR on our primers the prior week, we began the week with running a gel on the PCR products in addition to our most recently made plasmid. Our primers looked sub-optimal on our gel, but this was most likely due to an error in determining the annealing temperature for our reverse as opposed to bad primers. The plasmid however, looked quite bad. Despite it's nanodrop values being optimal, the sample on the gel looked quite dirty. Despite this, three transformations were conducted. These transformations, as expected, were unsuccessful. They appeared to show contamination. This raised concerns about our D. rad competent cells being possibly contaminated. We performed gram stains on these cells and they did not show any signs of contamination. This week we also observed our 3-way unable to grow onto 8µg/ml kanamycin plates from freezebac. We attempted to again inoculate onto antibiotic plates, having to wait until the following week to see results. After our plasmid showing improper on our gel, we performed three more plasmid extractions of our four-way E. coli. Our nanodrop readings showed to be quite ideal. We would run a gel of our extractions the following week to confirm their cleanliness.
Gel layoutfrom top to bottom: 1kb, 100bp, forward, forward, reverse, reverse, kan, plasmid

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