Gel shift assay: Demonstration of enhanced binding of oligo(δ)-L-ornithine-oligodeoxynucleotide conjugates to complementary DNA and RNA

Tianmin Zhu, Shahriar Pooyan, Ziping Wei, Michael J Leibowitz, Stanley Stein

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

An increase in melting temperature for DNA:DNA duplexes had been observed previously (Zhu et al. Antisense Res. Dev. 3:349-356, 1993) when an oligo(δ)ornithine moiety was covalently appended to a short oligodeoxynucleotide. We now report the analysis of duplex formation by electrophoretic gel shift analysis. In the particular example studied, an increase in Tm of 4°C was found to correspond to about a fivefold increase in binding constant. A similar enhancement by the appended cationic peptide was observed when the target strand was RNA. The use of a competitive assay format for avoidance of adsorptive loss at low concentrations (<10-7 M) of the oligonucleotide-oligo(δ)ornithine conjugate is presented.

Original languageEnglish (US)
Pages (from-to)69-74
Number of pages6
JournalAntisense and Nucleic Acid Drug Development
Volume6
Issue number1
StatePublished - Mar 1996
Externally publishedYes

ASJC Scopus subject areas

  • Genetics
  • Pharmacology

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