Objectives: Gene expression analysis by quantitative PCR is a standard laboratory technique for RNA quantification with high accuracy. In particular real-time PCR techniques using SYBR Green and melting curve analysis allowing verification of specific product amplification have become a well accepted laboratory technique for rapid and high throughput gene expression quantification. However, the software that is applied for quantification is somewhat circuitous and needs actually above average manual operation.
Design and methods: We here developed a novel, simple to handle open source software package (i.e., MAKERGAUL) for quantification of gene expression data obtained by real time PCR technology.
Results: The developed software was evaluated with an already well characterized real time PCR data set and the performance parameters (i.e., absolute bias, linearity, reproducibility, and resolution) of the algorithm that are the basis of our calculation procedure compared and ranked with those of other implemented and well-established algorithms. It shows good quantification performance with reduced requirements in computing power.
Conclusions: We conclude that MAKERGAUL is a convenient and easy to handle software allowing accurate and fast expression data analysis.
Keywords: CPU; Computer-based analysis; DNA quantification; Expression analysis; MAK2; MAKERGAUL; MIQE; Open source software; RT; central processing unit; double stranded DNA; dsDNA; qPCR; quantitative PCR; reverse transcription.