Cloning PCR Products By Addition Of Restriction Sites To The Termini Of Amplified DNA (Subscription Required) Protocol

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Protocol Information

Title Cloning PCR Products By Addition Of Restriction Sites To The Termini Of Amplified DNA (Subscription Required)
Source CSH Protocols; 2006; doi:10.1101/pdb.prot3835

Overview

Pairs of oligonucleotide primers used in PCR are often designed with restriction sites in their 5' regions. In many cases, the sites are different in the two primers. In this case, amplification generates a target fragment whose termini now carry new restriction sites that can be used for directional cloning into plasmid vectors. The purified fragment and the vector are digested with the appropriate restriction enzymes, ligated together, and transformed into E. coli. By Joseph Sambrook and David W. Russell.

Details

Details of this protocol, Cloning PCR Products By Addition Of Restriction Sites To The Termini Of Amplified DNA (Subscription Required), are located on a web site other than Biocompare Protocols.

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