Analyzing ligation mixtures using a PCR based method
- 1 January 2005
- journal article
- Published by Springer Science and Business Media LLC in Biological Procedures Online
- Vol. 7 (1), 93-100
- https://doi.org/10.1251/bpo108
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.Keywords
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