Title

A Single Molecular Beacon Probe Is Sufficient For The Analysis Of Multiple Nucleic Acid Sequences

Keywords

Biosensors; DNA recognition; Molecular beacons; Nucleic acids

Abstract

Molecular beacon (MB) probes are dual-labeled hairpin-shaped oligodeoxyribonucleotides that are extensively used for real-time detection of specific RNA/DNA analytes. In the MB probe, the loop fragment is complementary to the analyte: therefore, a unique probe is required for the analysis of each new analyte sequence. The conjugation of an oligonucleotide with two dyes and subsequent purification procedures add to the cost of MB probes, thus reducing their application in multiplex formats. Here we demonstrate how one MB probe can be used for the analysis of an arbitrary nucleic acid. The approach takes advantage of two oligonucleotide adaptor strands, each of which contains a fragment complementary to the analyte and a fragment complementary to an MB probe. The presence of the analyte leads to association of MB probe and the two DNA strands in quadripartite complex. The MB probe fluorescently reports the formation of this complex. In this design, the MB does not bind the analyte directly; therefore, the MB sequence is independent of the analyte. In this study one universal MB probe was used to genotype three human polymorphic sites. This approach promises to reduce the cost of multiplex real-time assays and improve the accuracy of single-nucleotide polymorphism genotyping. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA.

Publication Date

8-16-2010

Publication Title

ChemBioChem

Volume

11

Issue

12

Number of Pages

1762-1768

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1002/cbic.201000287

Socpus ID

77955618054 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/77955618054

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