A Simple Procedure for Cross-Linking Complementary Oligonucleotides

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Abstract
A simple, efficient procedure for cross-linking two complementary oligonucleotides, which does not require chemical modification of either oligonucleotide, is described. One of the oligonucleotides is first converted to the 5′-phosphorothioate derivative with polynucleotide kinase. It is then incubated with its complement in the presence of 1 μM trans-platinum(II)diammine dichloride. After overnight incubation, 40–50% cross-linking is observed. DNA synthesis by the Klenow fragment of Escherichia coli DNA polymerase I is blocked at the cross-linked site, resulting in the formation of truncated products. Potassium platinous chloride (K2PtCl4) and cis-platinum(II)diammine dichloride form cross-links less efficiently than the trans isomer.