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Quantitation of mRNA by Competitive PCR Using Capillary Electrophoresis

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 163))

Abstract

The high sensitivity of the reverse transcriptase-polymerase chain reaction (RT-PCR) allows the amplification of low abundance mRNA transcripts, but the biomedical research and diagnostics community have increasing interest in the ability to accurately quantify the amounts of mRNA in cell samples. Although theoretically it should be possible to calculate the starting copy number from a consideration of the efficiency of the reverse transcription step of the reaction and the number of PCR cycles performed, it is widely recognized that this approach is flawed and can lead to highly inaccurate results (1).

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References

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© 2001 Humana Press Inc.

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Williams, S.J., Williams, P.M. (2001). Quantitation of mRNA by Competitive PCR Using Capillary Electrophoresis. In: Mitchelson, K.R., Cheng, J. (eds) Capillary Electrophoresis of Nucleic Acids. Methods in Molecular Biology™, vol 163. Humana Press. https://doi.org/10.1385/1-59259-116-7:243

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  • DOI: https://doi.org/10.1385/1-59259-116-7:243

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-765-6

  • Online ISBN: 978-1-59259-116-9

  • eBook Packages: Springer Protocols

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