[Abstract]  [Full Text]  [PDF]

Prafulla K. Chandra^1* and Stephen K. Wikel¹

¹ Center for Microbial Pathogenesis, MC3710, School of Medicine, University of Connecticut Health Center. Farmington, CT. USA.

* To whom correspondence should be addressed: Prafulla K. Chandra, Center for Microbial Pathogenesis, MC3710, School of Medicine, University of Connecticut Health Center. 263 Farmington Avenue, Farmington, CT 06030-3710. USA. Email: pchandra@uchc.edu

Biol. Proced. Online 2005;7:93-100. doi:10.1251/bpo108
Submitted: March 07, 2005; Accepted: May 20, 2005; Published: June 17, 2005.

Indexing terms: Ligation; Nucleic Acids; Polymerase Chain Reaction.

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.