In fact, until the advent of the polymerase chain reaction (PCR) ( 4), they provided the most convenient way to manipulate individual genes and move them from one vector to another. As methods for the determination of DNA sequence improved, it became convenient to search those sequences for potential restriction enzyme cleavage sites since that would permit the facile further manipulation of specific fragments. In the 1970s they were used extensively to provide physical maps of small DNAs and, in the 1980s, were used to map large DNAs. A comprehensive database (REBASE) contains information about these enzymes including their recognition specificities and their sensitivity to DNA methylation ( 3). These enzymes recognize short DNA sequences (4–8 nucleotides) and cleave at, or close to, their recognition sites ( 1, 2). ![]() The Type II restriction enzymes are among the most valuable tools available to researchers in molecular biology.
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