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Abstract
A technique for conveniently radiolabeling DNA restriction endonuclease fragments
to high specific activity is described. DNA fragments are purified from agarose gels
directly by ethanol precipitation and are then denatured and labeled with the large
fragment of DNA polymerase I, using random oligonucleotides as primers. Over 70% of
the precursor triphosphate is routinely incorporated into complementary DNA, and specific
activities of over 10(9) dpm/microgram of DNA can be obtained using relatively small
amounts of precursor. These "oligolabeled" DNA fragments serve as efficient probes
in filter hybridization experiments.