Assessing and reducing sources of gene expression variability in Staphylococcus epidermidis biofilms. | - CCMAR -

Journal Article

TitleAssessing and reducing sources of gene expression variability in Staphylococcus epidermidis biofilms.
Publication TypeJournal Article
AuthorsSousa, C
Secondary AuthorsFrança, Â
Corporate AuthorsCerca, Nuno
Year of Publication2014
JournalBiotechniques
Volume57
Issue6
Date Published2014 Dec
Pagination295-301
ISSN1940-9818
KeywordsBioengineering, Biofilms, Biotechnology, DNA, Bacterial, Gene Expression, RNA, Bacterial, RNA, Messenger, Staphylococcus epidermidis
Abstract

Gene expression quantification can be a useful tool in studying the properties of bacterial biofilms. Unfortunately, techniques such as RNA extraction, cDNA synthesis, and quantitative PCR (qPCR) can introduce variability into mRNA transcript measurements, obscuring biologically relevant results. Here we sought to identify the steps that impair accurate gene expression quantification from Staphylococcus epidermidis biofilm samples. We devised an experimental setup that could be used to determine the contribution of each experimental step to the variability of mRNA transcript measurement. Among factors tested, biofilm growth contributed the most bias to gene expression quantification. Additional experiments demonstrated that pooling biofilms together reduced this variability, resulting in more accurate gene expression analysis results. We therefore recommend pooling in order to reduce the variability associated with gene expression quantification from biofilm samples.

DOI10.2144/000114238
Sapientia

http://www.ncbi.nlm.nih.gov/pubmed/25495729?dopt=Abstract

Alternate JournalBioTechniques
PubMed ID25495729
CCMAR Authors