A standard curve based method for relative real time PCR data processing

Background: Currently real time PCR is the most precise method by which to measure gene expression. The method generates a large amount of raw numerical data and processing may notably influence final results. The data processing is based either on standard curves or on PCR efficiency assessment. At the moment, the PCR efficiency approach is preferred in relative PCR whilst the standard curve is often used for absolute PCR. However, there are no barriers to employ standard curves for relative PCR. This article provides an implementation of the standard curve method and discusses its advantages and limitations in relative real time PCR. Results: We designed a procedure for data processing in relative real time PCR. The procedure completely avoids PCR efficiency assessment, minimizes operator involvement and provides a statistical assessment of intra-assay variation. The procedure includes the following steps. (I) Noise is filtered from raw fluorescence readings by smoothing, baseline s...

• Type: Journal
• Year: 2005
• Where: BMCBI
• Authors: Alexey Larionov, Andreas Krause, William Miller
• Keywords: BMCBI 2005, PCR Efficiency, PCR Efficiency Assessment, Standard Curve