840:153g:Projects/project27/2012/11/08

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Failed Transformation

  • The transformation from 11/6 showed no usable results. Colonies of positive control cells were present, grown on ampicillin media. Colonies were also present of negative control cells (transformed with water) and untransformed cells that were grown on media not containing antibiotics. These results indicate that the transformation was performed correctly, but that the target plasmid-promoter complex was not appropriately transformed into the competent cells
  • A PCR was performed according to the protocol designed on 11/6, using M. trichosporium genomic DNA as template, and both primer sets.
  • A ligation of the J23100 promoter and each of the two target vectors, pSB1K3 and J61002, was again attempted. This was performed at 4°C to test whether ligation temperature would increase the rate of successful ligation and therefore lead to the transformation of a functional plasmid-insert complex into E. coli.
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