PNA-Based Light-Up Probes for Real-Time Detection of Sequence-Specific PCR Products | Zendy

Petra Wolffs | Zendy; Rickard Knutsson | Zendy; Robert Sjöback | Zendy; Peter Rådström | Zendy

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PNA-Based Light-Up Probes for Real-Time Detection of Sequence-Specific PCR Products

Author(s) -

Petra Wolffs,

Rickard Knutsson,

Robert Sjöback,

Peter Rådström

Publication year - 2001

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/01314st01

Subject(s) - peptide nucleic acid , detection limit , yersinia enterocolitica , microbiology and biotechnology , dna , primer dimer , real time polymerase chain reaction , biology , genomic dna , nucleic acid , fluorescence , chemistry , polymerase chain reaction , chromatography , biochemistry , gene , genetics , bacteria , multiplex polymerase chain reaction , physics , quantum mechanics

The aim of this study was to introduce the use of a peptide nucleic acid (PNA)-thiazole orange conjugate for real-time monitoring of PCR. When the so-called light-up probes hybridize sequence-specifically to the PCR product, an increase in the fluorescent signal is obtained. It was found that the light-up probe can quantitatively measure the amount of DNA or intact bacterial cells in the reaction mixture, without interfering with the PCR amplification. A linear detection range of at least 4 log units was obtained without optimization of the system. The detection limit of this light-up assay per reaction mixture was 0.4 pg genomic Yersinia enterocolitica DNA.

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