A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity

✍ Scribed by Andrew P. Feinberg; Bert Vogelstein

• Book ID: 102984817
• Publisher: Elsevier Science
• Year: 1983
• Tongue: English
• Weight: 973 KB
• Volume: 132
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(83)90418-9

✦ Synopsis

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.