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Real-Time PCR and Multiplex Approaches

  • Olga L. GurvichEmail author
  • Mikhail Skoblov
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 784)

Abstract

Analysis of RNA expression levels by real-time reverse-transcription (RT) PCR has become a routine technique in diagnostic and research laboratories. Monitoring of DNA amplification can be done using fluorescent sequence-specific probes, which generate signal only upon binding to their target. Numerous fluorescent dyes with unique emission spectra are available and can be used to differentially label probes for various genes. Such probes can be added to the same PCR amplification reaction for simultaneous detection of multiple targets in a single assay. Such multiplexing is advantageous, since it markedly increases throughput and decreases costs and labor. Here, we describe application of multiplex real-time RT-PCR using TaqMan probes in the analysis of relative expression levels of a novel tumor-associated gene CUG2 in cell lines and tissue samples.

Key words

Multiplex Real-time PCR Gene expression TaqMan probes CUG2 C6orf173 

Notes

Acknowledgments

Authors would like to thank Alexander Skoblov and Michelle Nyhan for their help and advice in preparation of the manuscript.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Moscow State UniversityMoscowRussia
  2. 2.Research Centre for Medical GeneticsRussian Academy of Medical SciencesMoscowRussia

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