A Bayesian method for calculating real-time quantitative PCR calibration curves using absolute plasmid DNA standards

Background: In real-time quantitative PCR studies using absolute plasmid DNA standards, a calibration curve is developed to estimate an unknown DNA concentration. However, potential differences in the amplification performance of plasmid DNA compared to genomic DNA standards are often ignored in calibration calculations and in some cases impossible to characterize. A flexible statistical method that can account for uncertainty between plasmid and genomic DNA targets, replicate testing, and experiment-to-experiment variability is needed to estimate calibration curve parameters such as intercept and slope. Here we report the use of a Bayesian approach to generate calibration curves for the enumeration of target DNA from genomic DNA samples using absolute plasmid DNA standards. Results: Instead of the two traditional methods (classical and inverse), a Monte Carlo Markov Chain (MCMC) estimation was used to generate single, master, and modified calibration curves. The mean and the percenti...

• Type: Journal
• Year: 2008
• Where: BMCBI
• Authors: Mano Sivaganesan, Shawn Seifring, Manju Varma, Richard A. Haugland, Orin C. Shanks
• Keywords: Absolute Plasmid Dna, BMCBI 2008, Calibration Curves, Plasmid Dna Standards