Artículo
Model based analysis of real-time PCR data from DNA binding dye protocols
Alvarez, Mariano Javier; Vila Ortiz, Guillermo J.; Salibe, Mariano C.; Podhajcer, Osvaldo Luis
; Pitossi, Fernando Juan


Fecha de publicación:
03/2007
Editorial:
BioMed Central
Revista:
BMC Bioinformatics
ISSN:
1471-2105
e-ISSN:
1471-2105
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Background: Reverse transcription followed by real-time PCR is widely used for quantification of specific mRNA, and with the use of double-stranded DNA binding dyes it is becoming a standard for microarray data validation. Despite the kinetic information generated by real-time PCR, most popular analysis methods assume constant amplification efficiency among samples, introducing strong biases when amplification efficiencies are not the same. Results: We present here anew mathematical model based on the classic exponential description of the PCR, but modeling amplification efficiency as a sigmoidal function of the product yield. The model was validated with experimental results and used for the development of a new method for real-time PCR data analysis. This model based method for real-time PCR data analysis showed the best accuracy and precision compared with previous methods when used for quantification of in-silico generated and experimental real-time PCR results. Moreover, the method is suitable for the analyses of samples with similar or dissimilar amplification efficiency. Conclusion: The presented method showed the best accuracy and precision. Moreover, it does not depend on calibration curves, making it ideal for fully automated high-throughput applications.
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Articulos(IIBBA)
Articulos de INST.DE INVEST.BIOQUIMICAS DE BS.AS(I)
Articulos de INST.DE INVEST.BIOQUIMICAS DE BS.AS(I)
Citación
Alvarez, Mariano Javier; Vila Ortiz, Guillermo J.; Salibe, Mariano C.; Podhajcer, Osvaldo Luis; Pitossi, Fernando Juan; Model based analysis of real-time PCR data from DNA binding dye protocols; BioMed Central; BMC Bioinformatics; 8; 85; 3-2007; 1-10
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