Efficient cloning of PCR generated DNA containing terminal restriction endonuclease recognition sites.

@article{Jung1990EfficientCO,
  title={Efficient cloning of PCR generated DNA containing terminal restriction endonuclease recognition sites.},
  author={Volker Jung and Sidney Pestka},
  journal={Nucleic acids research},
  year={1990},
  volume={18 20},
  pages={6156}
}
DNA generated by the polymerase chain reaction (PCR) (1) containing terminal restriction endonuclease recognition sites to permit cloning usually relies on the use of unphosphorylated primers incorporating a restriction endonuclease recognition site of choice plus 3 —4 extra 5' bases flanking that site. Various sites (Notl, Xhol and Xbal (2), for example) incorporated into the termini of PCR products have proven difficult to cut with their respective restriction endonucleases. Possible… CONTINUE READING
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Experimental Conditions Control Proteinase K Klenow T4 DNA polymerase Spermidine Prolonged Digestion with Restriction Endonuclease Xhol 4 hr overnight Concatamerization Cloning Efficiency

  • Jung.V, Jones.C, Kumar.C.S, S. Stefanos, O'Connell.S, Pestka.S
  • 1990

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