Cross Priming Amplification: Mechanism and Optimization for Isothermal DNA Amplification

CPA is a class of isothermal amplification reactions that is carried out by a strand displacement DNA polymerase and does not require an initial denaturation step or the addition of a nicking enzyme. At the assay temperature of 63°C, the formation of a primer-template hybrid at transient, spontaneou...

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Main Authors:	Xu, Gaolian, Hu, Lin, Zhong, Huayan, Wang, Hongying, Yusa, Sei-ichi, Weiss, Tristen C., Romaniuk, Paul J., Pickerill, Sam, You, Qimin
Format:	Online
Language:	English
Published:	Nature Publishing Group 2012
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3271364/

id	pubmed-3271364
recordtype	oai_dc
spelling	pubmed-32713642012-02-03 Cross Priming Amplification: Mechanism and Optimization for Isothermal DNA Amplification Xu, Gaolian Hu, Lin Zhong, Huayan Wang, Hongying Yusa, Sei-ichi Weiss, Tristen C. Romaniuk, Paul J. Pickerill, Sam You, Qimin Article CPA is a class of isothermal amplification reactions that is carried out by a strand displacement DNA polymerase and does not require an initial denaturation step or the addition of a nicking enzyme. At the assay temperature of 63°C, the formation of a primer-template hybrid at transient, spontaneous denaturation bubbles in the DNA template is favored over re-annealing of the template strands by the high concentration of primer relative to template DNA. Strand displacement is encouraged by the annealing of cross primers with 5′ ends that are not complementary to the template strand and the binding of a displacement primer upstream of the crossing primer. The resulting exponential amplification of target DNA is highly specific and highly sensitive, producing amplicons from as few as four bacterial cells. Here we report on the basic CPA mechanism – single crossing CPA – and provide details on alternative mechanisms. Nature Publishing Group 2012-02-03 /pmc/articles/PMC3271364/ /pubmed/22355758 http://dx.doi.org/10.1038/srep00246 Text en Copyright © 2012, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-sa/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-ShareALike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Xu, Gaolian Hu, Lin Zhong, Huayan Wang, Hongying Yusa, Sei-ichi Weiss, Tristen C. Romaniuk, Paul J. Pickerill, Sam You, Qimin
spellingShingle	Xu, Gaolian Hu, Lin Zhong, Huayan Wang, Hongying Yusa, Sei-ichi Weiss, Tristen C. Romaniuk, Paul J. Pickerill, Sam You, Qimin Cross Priming Amplification: Mechanism and Optimization for Isothermal DNA Amplification
author_facet	Xu, Gaolian Hu, Lin Zhong, Huayan Wang, Hongying Yusa, Sei-ichi Weiss, Tristen C. Romaniuk, Paul J. Pickerill, Sam You, Qimin
author_sort	Xu, Gaolian
title	Cross Priming Amplification: Mechanism and Optimization for Isothermal DNA Amplification
title_short	Cross Priming Amplification: Mechanism and Optimization for Isothermal DNA Amplification
title_full	Cross Priming Amplification: Mechanism and Optimization for Isothermal DNA Amplification
title_fullStr	Cross Priming Amplification: Mechanism and Optimization for Isothermal DNA Amplification
title_full_unstemmed	Cross Priming Amplification: Mechanism and Optimization for Isothermal DNA Amplification
title_sort	cross priming amplification: mechanism and optimization for isothermal dna amplification
description	CPA is a class of isothermal amplification reactions that is carried out by a strand displacement DNA polymerase and does not require an initial denaturation step or the addition of a nicking enzyme. At the assay temperature of 63°C, the formation of a primer-template hybrid at transient, spontaneous denaturation bubbles in the DNA template is favored over re-annealing of the template strands by the high concentration of primer relative to template DNA. Strand displacement is encouraged by the annealing of cross primers with 5′ ends that are not complementary to the template strand and the binding of a displacement primer upstream of the crossing primer. The resulting exponential amplification of target DNA is highly specific and highly sensitive, producing amplicons from as few as four bacterial cells. Here we report on the basic CPA mechanism – single crossing CPA – and provide details on alternative mechanisms.
publisher	Nature Publishing Group
publishDate	2012
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3271364/
_version_	1611503536475996160

Cross Priming Amplification: Mechanism and Optimization for Isothermal DNA Amplification

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