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

Alexander Ulrich, Kasper Røjkjær Andersen, Thomas U Schwartz

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

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.

Original languageEnglish
JournalPLOS ONE
Volume7
Issue12
Pages (from-to)e53360
ISSN1932-6203
DOIs
Publication statusPublished - 2012
Externally publishedYes

Keywords

  • Cloning, Molecular
  • DNA
  • Genetic Vectors
  • Plasmids
  • Polymerase Chain Reaction

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