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Accuracy in amplification

Nature Biotechnology volume 19, pages 1011–1012 (2001)Cite this article

To the editor

Sensitive quantitative PCR techniques based on coamplification of multiple target templates1,2,3 as well as kinetic PCR4,5,6 have been used to quantify tiny amounts of viral or ectopic DNA or mRNA expression, even in single isolated cells. As kinetic PCR systems measure the absolute amounts of a single nucleic acid in a sample, many applications, such as accurate mRNA expression analysis, often require the use of a reference template coexpressed in the analyzed cells. In such cases, we contend that stochastic variation in low-copy-number target templates defines the accuracy of sensitive amplification techniques for quantifying relative template levels.

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References

  1. Wang, A.M., Doyle, M.V. & Mark, D.F. Proc. Natl. Acad. Sci. USA 86, 9717–9721 (1989).

    Article  CAS  Google Scholar 

  2. Becker-André, M. & Hahlbrock, K. Nucleic Acids Res. 17, 9437–9446 (1989).

    Article  Google Scholar 

  3. Gilliland, G., Perrin, S., Blanchard, K. & Bunn, H.F. Proc. Natl. Acad. Sci.USA 87, 2725–2729 (1990).

    Article  CAS  Google Scholar 

  4. Holland, P.M., Abramson, R.D., Watson, R. & Gelfand, D.H. Proc. Natl. Acad. Sci. USA 88, 7276–7280 (1991).

    Article  CAS  Google Scholar 

  5. Livak, K.J., Flood, S.J., Marmaro, J., Giusti, W. & Deetz, K. PCR Methods Appl. 4, 357–362 (1995).

    Article  CAS  Google Scholar 

  6. Wittwer, C.T., Herrmann, M.G., Moss, A.A. & Rasmussen, R.P. BioTechniques 22, 130–131, 134–138 (1997).

    Article  CAS  Google Scholar 

  7. Karrer, E.E. et al. Proc. Natl. Acad. Sci. USA 92, 3814–3818 (1995).

    Article  CAS  Google Scholar 

  8. Peccoud, J. & Jacob, C. Biophys. J. 71, 101–108 (1996).

    Article  CAS  Google Scholar 

  9. Sykes, P.J. et al. Biotechniques 13, 444–449 (1992).

    CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Clinical Chemistry, University of Helsinki and Laboratory, Helsinki University Central Hospital, Helsinki, Finland

    Jakob Stenman & Arto Orpana

  2. Department of Medical Genestics, University of Helsinki and Laboratory, Helsinki University Central Hospital, Helsinki, Finland

    Arto Orpana

Authors
  1. Jakob Stenman
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  2. Arto Orpana
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Stenman, J., Orpana, A. Accuracy in amplification. Nat Biotechnol 19, 1011–1012 (2001). https://doi.org/10.1038/nbt1101-1011b

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