Comparison of automated LRE quantification with LinReg and Miner via comparison to LDA quantification. Amplification profiles for each of the five enzyme formulations representative of all eleven transcripts (additional file 5) were subjected to analysis with: (A) Java-LRE analysis using the prototypic Java program that automates LRE analysis (additional file 4) (B) LinReg which determines amplification efficiency and F0 via linear regression using the FC readings within the log-linear region. (C) Miner which determines amplification efficiency using non-linear regression of the log-linear region, along with Ct values based upon a dynamic Ft. The resulting F0 values converted to the number of target molecules (N0) via optical calibration (see text). The quantities produced by each of the three methods are compared to the absolute quantification generated by limiting dilution assay (Table 3) via Log2 plots. Based on the resulting r2 values, LRE produced the highest quantitative accuracy for all five a formulations.
Rutledge and Stewart BMC Biotechnology 2008 8:47 doi:10.1186/1472-6750-8-47