Quantitative real-time polymerase chain reaction (qPCR) is a sensitive tool that can be used to quantify and compare the amount of specific RNA transcripts between different biological samples. This chapter describes the use of a “two-step” qPCR method to calculate the relative fold change of expression of genes of interest in S. aureus. Using this work-flow, cDNA is synthesized from RNA templates (previously checked for the absence of significant genomic DNA contamination) using a cocktail of random primers and reverse-transcriptase enzyme. The cDNA pools generated can then be assessed for expression of specific genes of interest using SYBR Green-based qPCR and quantification of relative fold-change expression.
CITATION STYLE
Lewis, A. M., & Rice, K. C. (2016). Quantitative real-time PCR (qPCR) workflow for analyzing staphylococcus aureus gene expression. In Methods in Molecular Biology (Vol. 1373, pp. 143–154). Humana Press Inc. https://doi.org/10.1007/7651_2014_193
Mendeley helps you to discover research relevant for your work.