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pPCV, a versatile vector for cloning PCR products

The efficiency of PCR product cloning depends on the nature of the DNA polymerase employed because amplicons may have blunt-ends or 3′ adenosines overhangs. Therefore, for amplicon cloning, available commercial vectors are either blunt-ended or have a single 3′ overhanging thymidine. The aim of this...

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Autores principales: Janner, Christiane R, Brito, Ana Lívia P, Moraes, Lidia Maria P, Reis, Viviane CB, Torres, Fernando AG
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3777018/
https://www.ncbi.nlm.nih.gov/pubmed/24058893
http://dx.doi.org/10.1186/2193-1801-2-441
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author Janner, Christiane R
Brito, Ana Lívia P
Moraes, Lidia Maria P
Reis, Viviane CB
Torres, Fernando AG
author_facet Janner, Christiane R
Brito, Ana Lívia P
Moraes, Lidia Maria P
Reis, Viviane CB
Torres, Fernando AG
author_sort Janner, Christiane R
collection PubMed
description The efficiency of PCR product cloning depends on the nature of the DNA polymerase employed because amplicons may have blunt-ends or 3′ adenosines overhangs. Therefore, for amplicon cloning, available commercial vectors are either blunt-ended or have a single 3′ overhanging thymidine. The aim of this work was to offer in a single vector the ability to clone both types of PCR products. For that purpose, a minimal polylinker was designed to include restriction sites for EcoRV and XcmI which enable direct cloning of amplicons bearing blunt-ends or A-overhangs, respectively, still offering blue/white selection. When tested, the resulting vector, pPCV, presented high efficiency cloning of both types of amplicons.
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spelling pubmed-37770182013-09-20 pPCV, a versatile vector for cloning PCR products Janner, Christiane R Brito, Ana Lívia P Moraes, Lidia Maria P Reis, Viviane CB Torres, Fernando AG Springerplus Research The efficiency of PCR product cloning depends on the nature of the DNA polymerase employed because amplicons may have blunt-ends or 3′ adenosines overhangs. Therefore, for amplicon cloning, available commercial vectors are either blunt-ended or have a single 3′ overhanging thymidine. The aim of this work was to offer in a single vector the ability to clone both types of PCR products. For that purpose, a minimal polylinker was designed to include restriction sites for EcoRV and XcmI which enable direct cloning of amplicons bearing blunt-ends or A-overhangs, respectively, still offering blue/white selection. When tested, the resulting vector, pPCV, presented high efficiency cloning of both types of amplicons. Springer International Publishing 2013-09-05 /pmc/articles/PMC3777018/ /pubmed/24058893 http://dx.doi.org/10.1186/2193-1801-2-441 Text en © Janner et al.; licensee Springer. 2013 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Janner, Christiane R
Brito, Ana Lívia P
Moraes, Lidia Maria P
Reis, Viviane CB
Torres, Fernando AG
pPCV, a versatile vector for cloning PCR products
title pPCV, a versatile vector for cloning PCR products
title_full pPCV, a versatile vector for cloning PCR products
title_fullStr pPCV, a versatile vector for cloning PCR products
title_full_unstemmed pPCV, a versatile vector for cloning PCR products
title_short pPCV, a versatile vector for cloning PCR products
title_sort ppcv, a versatile vector for cloning pcr products
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3777018/
https://www.ncbi.nlm.nih.gov/pubmed/24058893
http://dx.doi.org/10.1186/2193-1801-2-441
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