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Universal Restriction Site-Free Cloning Method Using Chimeric Primers
A universal restriction site-free cloning method has been developed to precisely insert a DNA fragment into a vector at any desired location without altering any nucleotide(s) in either the DNA fragment or the vector. The technique employs two pairs of chimeric primers, each containing a ribonucleotide. One pair of primers is used to amplify a target DNA fragment and another is used to prepare a linear vector. The ribonucleotide is used as a specific site for cleavage promoted by rare-earthdoi:10.2144/02323st02 pmid:11926173 fatcat:32tzk4whzzeyligsvhtuibrpwm