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Self-reporting PNA/DNA primers for PCR analysis. | LitMetric

Self-reporting PNA/DNA primers for PCR analysis.

Genome Res

Boston Probes, Bedford, Massachusetts 01730, USA.

Published: April 2001

We report a new fluorogenic method for sealed-tube PCR analysis using a quencher-labeled peptide nucleic acid (Q-PNA) probe. The Q-PNA hybridizes to a complementary tag sequence located at the 5' end of a 5' fluorophore-labeled oligonucleotide primer, quenching the primer's fluorescence. Incorporation of the primer into a doublestranded amplicon causes displacement of the Q-PNA such that the fluorescence of the sample is a direct indication of the amplicon concentration. The Q-PNA is able to quench multiple primers bearing distinct 5' fluorophores in a single reaction. We show realtime quantitative detection of a single-copy gene, K-ras, from human genomic DNA, as well as an endpoint multiplex assay for Chlamydia trachomatis and Neisseria gonorrhoeae targets. Because the Q-PNA may be used to quench any primer that contains the 5' tag sequence, it is possible to inexpensively adapt an existing primer set for use in a self-reporting fluorescent assay by including the tag sequence in one of the primers.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC311047PMC
http://dx.doi.org/10.1101/gr.170401DOI Listing

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