Model based analysis of real-time PCR data from DNA binding dye protocols

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 a new 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 insilico generated and experimental real-time PCR results. Moreover, the meth...

• Type: Journal
• Year: 2007
• Where: BMCBI
• Authors: Mariano J. Alvarez, Guillermo J. Vila-Ortiz, Mariano C. Salibe, Osvaldo L. Podhajcer, Fernando J. Pitossi
• Keywords: Amplification Efficiency, BMCBI 2007, Real-time Pcr, Real-time Pcr Data