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Rapid Partial Digestion of DNA Fragments by PCR with 5-methyl-dCTP
By Kwong-Kwok Wong, Ph.D.
Molecular Biosciences, Pacific Northwest National Laboratory, Richland, WA
In this report, a method is described for generation of a partial digestion pattern
for any fragment that can be amplified by PCR. Using a mixture of 5-methyl-dCTP (5mC) and
dCTP in PCR protects some of the restriction sites from digestion with
5-methyl-cytosine-sensitive restriction enzymes. After PCR amplification with 5mC,
restriction digestion of the modified PCR products with a 5-methyl-cytosine-sensitive
enzyme rapidly produces an array of fragments similar to a partial digestion of
unmethylated DNA.
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