Blunt-Ended Cloning Into Plasmid Vectors (Subscripiton Required) Protocol

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Protocol Information

Title Blunt-Ended Cloning Into Plasmid Vectors (Subscripiton Required)
Source CSH Protocols; 2006; doi:10.1101/pdb.prot3921

Overview

Target DNA is ligated to a blunt-ended plasmid DNA, and the products of the ligation reaction are used to transform competent E. coli. The maximum number of "correct" clones can generally be obtained from ligation reactions containing equimolar amounts of plasmid and target DNAs, with the total DNA concentration being <100 µg/ml. Blunt-end ligation catalyzed by bacteriophage T4 DNA ligase is suppressed by high concentrations (5 mM) of ATP and polyamines such as spermidine. By Joseph Sambrook and David W. Russell.

Details

Details of this protocol, Blunt-Ended Cloning Into Plasmid Vectors (Subscripiton Required), are located on a web site other than Biocompare Protocols.

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