One-step generation of recombineering constructs by asymmetric-end ligation and negative selection.

  title={One-step generation of recombineering constructs by asymmetric-end ligation and negative selection.},
  author={M. Ohtsuka and A. Mizutani and Y. Y. Kikuti and J. Kulski and M. Sato and M. Kimura and M. Tanaka and H. Inoko},
  journal={Analytical biochemistry},
  volume={360 2},
Recombineering [1] or Red/ET recombination [2], a recently developed technique based on homologous recombination in Escherichia coli, is useful for the efficient production of a gene-targeting vector (TV) by modification of bacterial artificial chromosome (BAC) clones. This method requires the preparation of a linear DNA recombineering fragment, containing a selectable gene flanked by homologous regions (HRs) for recombination. The PCR method using primers containing HR sequences is the… Expand
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