Gene Expression Quantification from Pathogenic Bacterial Biofilms by Quantitative PCR

Abstract

Quantitative PCR (qPCR) is one of the most used techniques to quantify gene expression in bacterial biofilms due to its easiness, sensitivity, and robustness. However, several practical aspects need to be considered to obtain accurate and reliable results. Here, we describe a detailed and optimized protocol to quantify mRNA transcripts from bacterial biofilms using qPCR, including pieces of advice to improve RNA quality, which ultimately increases the accuracy, consistency, and relevance of gene expression data.

Keywords: Bacteria; Biofilms; Complementary DNA synthesis; DNase treatment; RNA isolation; RNA quality; Sample collection; qPCR.