A simple procedure for cross-linking complementary oligonucleotides.

B C Chu; L E Orgel

doi:10.1089/dna.1990.9.71

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A simple procedure for cross-linking complementary oligonucleotides.

B C Chu·L E Orgel

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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 microM 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.

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Base Sequence Cisplatin Cross-Linking Reagents Genetic Techniques Molecular Sequence Data Nucleic Acid Hybridization Oligodeoxyribonucleotides Oligonucleotides Polynucleotide 5'-Hydroxyl-Kinase Thionucleotides Non-programmatic Journal Article Research Support, U.S. Gov't, P.H.S.

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B C Chu, L E Orgel. A simple procedure for cross-linking complementary oligonucleotides.. https://doi.org/10.1089/dna.1990.9.71

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