Screening for oligonucleotide binding affinity by a convenient fluorescence competition assay.

J G Harrison; X Liu; S Balasubramanian

doi:10.1093/nar/27.17.e14

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Screening for oligonucleotide binding affinity by a convenient fluorescence competition assay.

J G Harrison·X Liu·S Balasubramanian

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Abstract

A competitive homogeneous quenched fluorescence assay system is described for the high throughput screening of DNA conjugates that bind to single-stranded DNA. Fluorescence signal is generated by competitive binding of the sample molecule to a target strand labelled with a quencher probe, which is otherwise hybridised to a complementary strand containing a fluorescent probe. Thus fluorescence generated is related to the affinity of the sample. Competitive analysis of a number of peptide-oligonucleotide conjugates gave data that correlated well with the corresponding UV melting data. The assay will be useful for screening of large numbers of potential single-stranded binding molecules.

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Chromatography, High Pressure Liquid DNA DNA, Single-Stranded Fluorescent Dyes Mass Spectrometry Nucleic Acid Denaturation Nucleic Acid Hybridization Spectrophotometry, Ultraviolet Temperature Journal Article Research Support, Non-U.S. Gov't

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J G Harrison, X Liu, S Balasubramanian. 1999-09-01. Screening for oligonucleotide binding affinity by a convenient fluorescence competition assay.. https://doi.org/10.1093/nar%2F27.17.e14

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