Cargando…
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...
Autores principales: | , , , , |
---|---|
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 |
_version_ | 1782284929138163712 |
---|---|
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. |
format | Online Article Text |
id | pubmed-3777018 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Springer International Publishing |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT jannerchristianer ppcvaversatilevectorforcloningpcrproducts AT britoanaliviap ppcvaversatilevectorforcloningpcrproducts AT moraeslidiamariap ppcvaversatilevectorforcloningpcrproducts AT reisvivianecb ppcvaversatilevectorforcloningpcrproducts AT torresfernandoag ppcvaversatilevectorforcloningpcrproducts |