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From 2011.igem.org

Short version: We tried transforming several different samples from two different mutagenesis pcr's: 6/30/11 and 7/14/11. For a positive control we used the sample 4A4 of undiluted pET-Taq that Dr Burkett supplied. Fot negative control we simply subjected the cells to the heat shock, etc. We also attempted to push the 5I and 5K samples into the NEB competent cells. No growth on any plates after about 22 hrs.

One conjecture: the cells were so cold when we heat-shocked them that they were not at the required 42°C for long enough time. I suggest we use the MIT- recommended 60 sec heat-shock.

We also selected 4 colonies from the plate that Dr Burkett gave us, which was labeled as having the 4A4 E coli cells (pET-Taq - E coli cloned with the Taq-pol1 gene minus one bp in middle), but these colonies also apparently showed no growth in Amp-laced LB. Hmmmmm