A homogeneous nucleic acid hybridization assay based on strand displacement.

Abstract:

:A homogeneous nucleic acid hybridization assay which is conducted in solution and requires no separation steps is described. The assay is based on the concept of strand displacement. In the strand displacement assay, an RNA "signal strand" is hybridized within a larger DNA strand termed the "probe strand", which is, in turn, complementary to the target nucleic acid of interest. Hybridization of the target nucleic acid with the probe strand ultimately results in displacement of the RNA signal strand. Strand displacement, therefore, causes conversion of the RNA from double to single-stranded form. The single-strand specificity of polynucleotide phosphorylase (EC 2.7.7.8) allows discrimination between double-helical and single-stranded forms of the RNA signal strand. As displacement proceeds, free RNA signal strands are preferentially phosphorolyzed to component nucleoside diphosphates, including adenosine diphosphate. The latter nucleotide is converted to ATP by pyruvate kinase(EC 2.7.1.40). Luciferase catalyzed bioluminescence is employed to measure the ATP generated as a result of strand displacement.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Vary CP

doi

10.1093/nar/15.17.6883

subject

Has Abstract

pub_date

1987-09-11 00:00:00

pages

6883-97

issue

17

eissn

0305-1048

issn

1362-4962

journal_volume

15

pub_type

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