| Literature DB >> 22328425 |
Mamie Z Li1, Stephen J Elledge.
Abstract
We describe here a method for sequence- and ligation-independent cloning (SLIC). SLIC uses an exonuclease, T4 DNA polymerase, to generate single-stranded DNA overhangs in insert and vector sequences. These fragments are then assembled in vitro and transformed into Escherichia coli to generate recombinant DNA of interest. SLIC inserts can also be generated by incomplete PCR (iPCR) or mixed PCR. As many as five inserts can be assembled in one reaction simultaneously with great efficiency using SLIC. SLIC circumvents sequence constraints for recombinant DNA using standard restriction enzyme-mediated cloning and previous ligation-independent cloning methods and provides a new approach for the efficient generation of recombinant DNA.Entities:
Mesh：
Substances：
Year: 2012 PMID： 22328425 DOI： 10.1007/978-1-61779-564-0_5
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745