Direct Preparation of Sticky-Ended Duplexes within PCR by Using Caged Primers

Abstract Control of the terminal structures of PCR products is crucially important to facilitate molecular biology and biotechnology. Here, we report a new method to prepare PCR products having desired sticky ends directly after thermal cycles. When a pair of caged primers is used, polymerase reacti...

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Bibliographic Details
Published in:Nucleic Acids Symposium Series 2008, Vol.52 (1), p.467-468
Main Authors: Tanaka, Keita, Katada, Hitoshi, Shigi, Narumi, Kuzuya, Akinori, Komiyama, Makoto
Format: Article
Language:English
Online Access:Request full text
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Summary:Abstract Control of the terminal structures of PCR products is crucially important to facilitate molecular biology and biotechnology. Here, we report a new method to prepare PCR products having desired sticky ends directly after thermal cycles. When a pair of caged primers is used, polymerase reaction is site-selectively terminated in front of the caged nucleotide, and the 5′- portion of the primer remains single-stranded throughout the reaction. Successive removal of the photo-cleavable protecting group gives restriction-enzyme free sticky ends on the product. We succeeded in applying this technique to make a recombinant vector bearing GFP gene.
ISSN:0261-3166
1746-8272
DOI:10.1093/nass/nrn237