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Analyzing ligation mixtures using a PCR based method

Abstract

We have developed a simple and effective method (Lig-PCR) for monitoring ligation reactions using PCR and primers that are common to many cloning vectors. Ligation mixtures can directly be used as templates and the results can be analyzed by conventional gel electrophoresis. The PCR products are representative of the recombinant molecules created during ligation and the corresponding transformants. Orientation of inserts can also be determined using an internal primer. The usefulness of this method has been demonstrated using ligation mixtures of two cDNA’s derived from the salivary glands of Aedes aegypti mosquitoes. The method described here is sensitive and easy to perform compared to currently available methods.

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Correspondence to Prafulla K. Chandra.

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Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/2.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Chandra, P.K., Wikel, S.K. Analyzing ligation mixtures using a PCR based method. Biol. Proced. Online 7, 93–100 (2005). https://doi.org/10.1251/bpo108

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  • DOI: https://doi.org/10.1251/bpo108

Indexing terms

  • Ligation
  • Nucleic Acids
  • Polymerase Chain Reaction