Plasmid construction by SLIC or sequence and ligation-independent cloning

Ryan E Hill; Julian J Eaton-Rye

doi:10.1007/978-1-62703-764-8_2

Plasmid construction by SLIC or sequence and ligation-independent cloning

Methods Mol Biol. 2014:1116:25-36. doi: 10.1007/978-1-62703-764-8_2.

Authors

Ryan E Hill¹, Julian J Eaton-Rye

Affiliation

¹ Department of Biochemistry, University of Otago, Dunedin, New Zealand.

PMID: 24395354
DOI: 10.1007/978-1-62703-764-8_2

Abstract

Sequence and ligation-independent cloning (Nat Methods 4:251-256, 2007) is a powerful tool for the construction of multi-fragment complex plasmids in a simple and efficient manner. Plasmids consisting of 6-7 DNA fragments can be assembled in a single day, with additional 2 days for screening and extraction. SLIC requires PCR products with overlapping regions of 30-40 bp at the 5' and 3' ends, T4 DNA polymerase, and an optional RecA protein for construction.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cloning, Molecular / methods*
DNA Primers / genetics
DNA Primers / metabolism
DNA-Directed DNA Polymerase / metabolism
Plasmids* / genetics
Plasmids* / metabolism
Polymerase Chain Reaction

Substances

DNA Primers
DNA-Directed DNA Polymerase