Exponential megapriming PCR (EMP) cloning-seamless DNA insertion into any target plasmid without sequence constraints

Alexander Ulrich; Kasper Røjkjær Andersen; Thomas U Schwartz

doi:10.1371/journal.pone.0053360

Exponential megapriming PCR (EMP) cloning-seamless DNA insertion into any target plasmid without sequence constraints

Alexander Ulrich, Kasper Røjkjær Andersen, Thomas U Schwartz

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

61 Citationer (Scopus)

Abstract

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.

Originalsprog	Engelsk
Tidsskrift	PLOS ONE
Vol/bind	7
Nummer	12
Sider (fra-til)	e53360
ISSN	1932-6203
DOI	https://doi.org/10.1371/journal.pone.0053360
Status	Udgivet - 2012
Udgivet eksternt	Ja

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10.1371/journal.pone.0053360

Citationsformater

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abstract = "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.",

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AU - Ulrich, Alexander

AU - Andersen, Kasper Røjkjær

AU - Schwartz, Thomas U

PY - 2012

Y1 - 2012

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AB - 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.

KW - Cloning, Molecular

KW - DNA

KW - Genetic Vectors

KW - Plasmids

KW - Polymerase Chain Reaction

U2 - 10.1371/journal.pone.0053360

DO - 10.1371/journal.pone.0053360

M3 - Journal article

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SN - 1932-6203

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SP - e53360

JO - PLOS ONE

JF - PLOS ONE

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Exponential megapriming PCR (EMP) cloning-seamless DNA insertion into any target plasmid without sequence constraints

Abstract

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