User:Eduardo Vladimir Munoz/Notebook/UNAM Genomics Mexico 2011/2011/08/26

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High Stability Plasmid Development

Abstract

  • Transformation of the ligations of DD repC-PCR E,S and DD B0015-pSB1AK3 E,X.
  • Transformation of the ligations of DD B0015-PCR E,P and DD pSB1T3 E,P.

Transformations

I followed our transformation protocol for both ligations and plated the resulting cells. The two ligations of pSB1AK3 were plated in Ampicilin LB, and the two ligations of pSB1T3 were plated in Tetracyclin.

No bacteria were found in the Tet plates, indicating that either the ligations or the transformations were defective.

Bacteria were found in the Amp plates. However it is necessary to move these bacteria to Kanamicin media, to avoid Amp resistants lacking the transformation.


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