title: Exponential megapriming PCR (EMP) cloning--seamless DNA insertion into any target plasmid without sequence constraints.
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.
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Alexander
authors.initials: A
authors.lastname: Ulrich
authors.forename: Kasper R
authors.initials: KR
authors.lastname: Andersen
authors.forename: Thomas U
authors.initials: TU
authors.lastname: Schwartz
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics*; D005822:Genetic Vectors; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004247:DNA