Exponential Megapriming PCR (EMP) Cloning—Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints

We present a fast, reliable and inexpensive restriction-free cloning method for seamless DNA insertion into any plasmid without sequence limitation. Exponential megapriming PCR (EMP) cloning requires two consecutive PCR steps and can be carried out in one day. We show that EMP cloning has a higher e...

Descripción completa

Detalles Bibliográficos
Autores principales: Ulrich, Alexander, Andersen, Kasper R., Schwartz, Thomas U.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3534072/
https://www.ncbi.nlm.nih.gov/pubmed/23300917
http://dx.doi.org/10.1371/journal.pone.0053360
_version_ 1782475265389101056
author Ulrich, Alexander
Andersen, Kasper R.
Schwartz, Thomas U.
author_facet Ulrich, Alexander
Andersen, Kasper R.
Schwartz, Thomas U.
author_sort Ulrich, Alexander
collection PubMed
description We present a fast, reliable and inexpensive restriction-free cloning method for seamless DNA insertion into any plasmid without sequence limitation. Exponential megapriming PCR (EMP) cloning requires two consecutive PCR steps and can be carried out in one day. We show that EMP cloning has a higher efficiency than restriction-free (RF) cloning, especially for long inserts above 2.5 kb. EMP further enables simultaneous cloning of multiple inserts.
format Online
Article
Text
id pubmed-3534072
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-35340722013-01-08 Exponential Megapriming PCR (EMP) Cloning—Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints Ulrich, Alexander Andersen, Kasper R. Schwartz, Thomas U. PLoS One Research Article We present a fast, reliable and inexpensive restriction-free cloning method for seamless DNA insertion into any plasmid without sequence limitation. Exponential megapriming PCR (EMP) cloning requires two consecutive PCR steps and can be carried out in one day. We show that EMP cloning has a higher efficiency than restriction-free (RF) cloning, especially for long inserts above 2.5 kb. EMP further enables simultaneous cloning of multiple inserts. Public Library of Science 2012-12-31 /pmc/articles/PMC3534072/ /pubmed/23300917 http://dx.doi.org/10.1371/journal.pone.0053360 Text en © 2012 Ulrich et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Ulrich, Alexander
Andersen, Kasper R.
Schwartz, Thomas U.
Exponential Megapriming PCR (EMP) Cloning—Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints
title Exponential Megapriming PCR (EMP) Cloning—Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints
title_full Exponential Megapriming PCR (EMP) Cloning—Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints
title_fullStr Exponential Megapriming PCR (EMP) Cloning—Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints
title_full_unstemmed Exponential Megapriming PCR (EMP) Cloning—Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints
title_short Exponential Megapriming PCR (EMP) Cloning—Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints
title_sort exponential megapriming pcr (emp) cloning—seamless dna insertion into any target plasmid without sequence constraints
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3534072/
https://www.ncbi.nlm.nih.gov/pubmed/23300917
http://dx.doi.org/10.1371/journal.pone.0053360
work_keys_str_mv AT ulrichalexander exponentialmegaprimingpcrempcloningseamlessdnainsertionintoanytargetplasmidwithoutsequenceconstraints
AT andersenkasperr exponentialmegaprimingpcrempcloningseamlessdnainsertionintoanytargetplasmidwithoutsequenceconstraints
AT schwartzthomasu exponentialmegaprimingpcrempcloningseamlessdnainsertionintoanytargetplasmidwithoutsequenceconstraints

Ejemplares similares