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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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
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creator Tanaka, Keita
Katada, Hitoshi
Shigi, Narumi
Kuzuya, Akinori
Komiyama, Makoto
description 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.
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title Direct Preparation of Sticky-Ended Duplexes within PCR by Using Caged Primers
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