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

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  • Alexander Ulrich, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, Institute für Chemie und Biochemie, Freie Universität Berlin, USA
  • Kasper Røjkjær Andersen
  • Thomas U Schwartz, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

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.

OriginalsprogEngelsk
TidsskriftPLOS ONE
Vol/bind7
Nummer12
Sider (fra-til)e53360
ISSN1932-6203
DOI
StatusUdgivet - 2012
Eksternt udgivetJa

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