A novel universal real-time PCR system using the attached universal duplex probes for quantitative analysis of nucleic acids

Litao Yang,Dabing Zhang,Wanqi Liang

doi:10.1016/j.jbiotec.2008.07.530

Litao Yang, Dabing Zhang + Show 1 more

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https://doi.org/10.1016/j.jbiotec.2008.07.530

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Abstract

A novel universal real-time PCR system using the attached universal duplex probes for quantitative analysis of nucleic acids

Highlights

Real-time PCR techniques are being widely used for nucleic acids analysis, but one limitation of current frequently employed real-time PCR is the high cost of the labeled probe for each target molecule
PCR is a powerful tool for the amplification of minimal amounts of initial target genetic sequences [1,2], and it has been widely used for quantitative analysis of nucleic acids in medical diagnostics, drug discovery, virus detection, environmental monitoring, gene expression analysis and the detection of genetically modification organisms (GMO) [3,4,5]
A complete attached universal duplex probes (AUDP) PCR assay is composed of a target DNA-specific primer pair, with one primer containing an attached universal template (UT) sequence, and a set of universal duplex probes containing both fluorescent probe (FP) and quenching probe (QP) (Figure 1A)

Summary

Introduction

Real-time PCR techniques are being widely used for nucleic acids analysis, but one limitation of current frequently employed real-time PCR is the high cost of the labeled probe for each target molecule. When the donor fluorophore and acceptor fluorophore are separated in the reaction by cleavage, the change in secondary structure or strand displacement of the fluorescent probe will result in increased fluorescence signal [23,24,25]. In these real-time PCR assays, either duallabeled probes or primers have to be synthesized, which limits the wide application of these methods due to incomplete quenching, complicated synthesis and labeling, and high costs [26]

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Discussion

Conclusion