LIBRO-Laboratório de Investigação em Biofilmes Rosário Oliveira, Centre of Biological Engineering, University of Minho, Braga, Portugal.
LABBELS-Associate Laboratory, Braga/Guimarães, Portugal.
Methods Mol Biol. 2023;2967:133-149. doi: 10.1007/978-1-0716-3358-8_11.
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.
实时荧光定量 PCR(qPCR)是一种用于定量细菌生物膜中基因表达的常用技术,因为它具有简单、灵敏和稳健的特点。然而,为了获得准确可靠的结果,需要考虑几个实际方面。在这里,我们描述了一种使用 qPCR 定量细菌生物膜中 mRNA 转录本的详细和优化的方案,包括一些建议,以提高 RNA 质量,从而最终提高基因表达数据的准确性、一致性和相关性。