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Analyzing ligation mixtures using a PCR based method

External protocol Created on 09 Apr 2014

Authors

Prafulla K. Chandra, Stephen K. Wikel

Summary

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 aegyptimosquitoes. The method described here is sensitive and easy to perform compared to currently available methods.

Further details

The protocol was imported from Springer Protocols. To see the entire protocol, click on the source link, or download the pdf here.

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