Abstract

Droplet digital PCR data-analysis: Issues and solutions

Olivier Thas, Professor in Biostatistics & Honorary Professor, Ghent University

The use of digital PCR for quantification of nucleic acids is rapidly growing. A major drawback remains the lack of flexible data analysis tools. I will demonstrate some of the important issues related to sources of variability and bias, and the setting of the threshold on the fluorescent signal. Next I show how publically available software tools can be used for absolute quantification, copy number variation, and relative gene expression. The data-analysis pipeline also includes a method for the selection of stable reference genes. Finally I will demonstrate a tool for power and sample size calculation for differential expression studies. All tools are freely available from a single website.