Improving Lambda Red Genome Engineering in Escherichia coli via Rational Removal of Endogenous Nucleases

@inproceedings{Mosberg2012ImprovingLR,
  title={Improving Lambda Red Genome Engineering in Escherichia coli via Rational Removal of Endogenous Nucleases},
  author={Joshua A W Mosberg and Christopher J. Gregg and Marc J. Lajoie and Harris H. Wang and George M. Church},
  booktitle={PloS one},
  year={2012}
}
Lambda Red recombineering is a powerful technique for making targeted genetic changes in bacteria. However, many applications are limited by the frequency of recombination. Previous studies have suggested that endogenous nucleases may hinder recombination by degrading the exogenous DNA used for recombineering. In this work, we identify ExoVII as a nuclease which degrades the ends of single-stranded DNA (ssDNA) oligonucleotides and double-stranded DNA (dsDNA) cassettes. Removing this nuclease… CONTINUE READING

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