A simple procedure for cross-linking complementary oligonucleotides.

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.

journal_name

DNA Cell Biol

journal_title

DNA and cell biology

authors

Chu BC,Orgel LE

doi

10.1089/dna.1990.9.71

subject

Has Abstract

pub_date

1990-01-01 00:00:00

pages

71-6

issue

1

eissn

1044-5498

issn

1557-7430

journal_volume

9

pub_type

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