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

PLoS One. 2012;7(12):e53360. doi: 10.1371/journal.pone.0053360. Epub 2012 Dec 31.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular / methods*
  • DNA / genetics*
  • Genetic Vectors
  • Plasmids
  • Polymerase Chain Reaction / methods*

Substances

  • DNA