Overview
This protocol is for directional blunt-end cloning of DNA fragments. The target DNA is PCR-amplified, 3'-extensions are polished with Pfu DNA polymerase, and the amplicon is ligated to a blunt-ended plasmid DNA. The products of the ligation reaction are used to transform competent Escherichia coli. A restriction enzyme is added to the ligation reaction (Liu and Schwartz 1992) to relinearize any self-religating vector DNA. By Gina L. Costa and Michael P. Weiner.
Details
Details of this protocol, Directional Cloning Of PCR Products (Subscription Required), are located on a web site other than Biocompare Protocols.