Influence of reagents formulation on real-time PCR parameters

  1. Burgos, J.S. 1
  2. Ramı́rez, C. 1
  3. Tenorio, R. 1
  4. Sastre, I. 1
  5. Bullido, M.J. 1
  1. 1 Centro de Biología Molecular Severo Ochoa
    info

    Centro de Biología Molecular Severo Ochoa

    Madrid, España

    ROR https://ror.org/03v9e8t09

Revista:
Molecular and Cellular Probes

ISSN: 0890-8508

Año de publicación: 2002

Volumen: 16

Número: 4

Páginas: 257-260

Tipo: Artículo

DOI: 10.1006/MCPR.2002.0419 GOOGLE SCHOLAR lock_openAcceso abierto editor

Otras publicaciones en: Molecular and Cellular Probes

Resumen

Real-time polymerase chain reaction (PCR) techniques are increasingly used to quantify target sequences for diagnostic and research purposes. Due to its ‘quantitative’ character, it is very important to determine the variability of this technique correlating with several experimental conditions. The objective of this study was to analyse the effect of manufacturing lots of PCR reagents on two main PCR parameters, specificity and sensitivity. For this study, we used four different amplicons, using either mouse genomic DNA or viral DNA. Although a PCR product could be obtained in any of the conditions, we observed that there are relevant variations in sensitivity depending on the reagents formulation. We conclude that different lots of reagents may determine the analytical performance of PCR assays indicating that reagents testing are of special importance when the PCR protocol is used for quantitative purposes.

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