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Quantification on the LightCycler

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Abstract

Over the last 15 years, PCR has become an essential part of most laboratories involved in biomedical research. PCR amplification turns a few attograms of a specific fragment of nucleic acid (far too little to be analyzed directly or used in biochemical reactions) into as much as a microgram of DNA.

Keywords

  • Human Immunodeficiency Virus Type
  • Fluorescence Resonance Energy Transfer
  • Noise Band
  • Amplification Curve
  • Threshold Line

These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Ferre F, Marchese A, Pezzoli P, Griffith S, Buxton E, Boyer V (1994) Quantative PCR: an overview. In: Mullis KB, Ferre F, Gibbs RA (eds) The polymerase chain reaction. Birkhauser, Boston 67–88

    Google Scholar 

  2. Higuchi R, Dollinger G, Walsh PS, Griffith R (1992) Simultaneous amplification and detection of specific DNA sequences. Biotechnology 10: 413–417

    PubMed  CrossRef  CAS  Google Scholar 

  3. Schuurman R, Descamps S, Weverling G, Kaye S, Tijnagel J, Williams I, van Leeuwen R, Tedder R, Boucher C, Brun-Vezinet F, Loveday C (1996) Multicenter comparison of three commercial methods of quantification of human immunodeficiency virus type 1 RNA in plasma. J Clin Microbiol 34: 3016–3022

    PubMed  CAS  Google Scholar 

  4. Hayward AL, Oefner PJ, Sabatini S, Kainer DB, Hinojos CA, Doris PA (1998) Modeling and analysis of competitive RT-PCR. Nucleic Acids Res 26 (11): 2511–8

    PubMed  CrossRef  CAS  Google Scholar 

  5. Wang Z, Spandoro J (1998) Determination of target copy number of quantitative standards used in PCR-based diagnostic assays. In: Ferre F Gene quantification. Birkhauser, Boston, pp 31–43

    Google Scholar 

  6. Morrison TB, Weis JJ, Wittwer CT (1998) Quantification of low copy transcripts by continuous SYBR Green I monitoring during amplification. Biotechniques 24: 954–958

    PubMed  CAS  Google Scholar 

  7. Kellogg DE, Rybalkin I, Chen S, Mukhamedova N, Vlasik T, Siebert PD, Chenchik A (1994) Taqstart antibody:“hot start” PCR facilitated by a neutralizing monoclonal directed against Taq DNA polymerase. Biotechniques June 16 (6): 1134–1137

    CAS  Google Scholar 

  8. Ririe KM, Rasmussen RP, Wittwer CT (1997) Product differentiation by analysis of DNA melting curves during the polymerase chain reaction. Anal Biochem 245: 154–160

    PubMed  CrossRef  CAS  Google Scholar 

  9. Rasmussen RP, Morrison TB, Herrmann MG, WittwerCT (1998) Quantitative PCR by continuous flourescence monitoring of a double strand DNA specific binding dye. BioChemica 8–11

    Google Scholar 

  10. Wittwer CT, Herrmann MG, Moss AA, Rasmussen RP (1997) Continuous flourescence monitoring of rapid cycle DNA amplification. Biotechniques 22: 130–138

    PubMed  CAS  Google Scholar 

  11. Holland PM, Abramson RD, Watson R, Gelfland DH (1991) Detection of specific polymerase chain reaction product by utilizing the 5’ to 3’ exonuclease activity of thermus aquaticius DNA polymerase. Proceedings of the National Academy of Science 88: 7276–80

    CrossRef  CAS  Google Scholar 

  12. Wittwer CT, Ririe KM, Rasmussen RP (1998) Flourescence monitoring of rapid cycle PCR for quantification. In: Ferre F (ed) Gene quantification. Birkhauser, Boston

    Google Scholar 

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© 2001 Springer-Verlag Berlin Heidelberg

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Rasmussen, R. (2001). Quantification on the LightCycler. In: Meuer, S., Wittwer, C., Nakagawara, KI. (eds) Rapid Cycle Real-Time PCR. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-59524-0_3

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  • DOI: https://doi.org/10.1007/978-3-642-59524-0_3

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-540-66736-0

  • Online ISBN: 978-3-642-59524-0

  • eBook Packages: Springer Book Archive