Open AccessScreening for oligonucleotide binding affinity by a convenient fluorescence competition assay
Author(s) -
Joseph G. Harrison,
Xueliang Liu,
Shankar Balasubramanian
Publication year - 1999
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/27.17.e14-i
Subject(s) - oligonucleotide , fluorescence , biology , dna , conjugate , competitive binding , high throughput screening , hybridization probe , ligand binding assay , binding site , biophysics , microbiology and biotechnology , combinatorial chemistry , biochemistry , receptor , chemistry , quantum mechanics , physics , mathematical analysis , mathematics
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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