Please use this identifier to cite or link to this item: http://hdl.handle.net/1822/31922

TitleAssessing and reducing sources of gene expression variability in Staphylococcus epidermidis biofilms
Author(s)Sousa, Cármen
França, Ângela
Cerca, Nuno
Keywordsbiofilms
gene expression variability
biofilm pool
RNA extraction
reverse transcriptase
qPCR
Issue dateDec-2014
PublisherEaton Publishing Company
JournalBioTechniques
CitationSousa, Cármen; França, Ângela; Cerca, Nuno, Assessing and reducing sources of gene expression variability in S. epidermidis biofilms. BioTechniques, 57(6), 295-301, 2014
Abstract(s)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.
TypeArticle
URIhttp://hdl.handle.net/1822/31922
DOI10.2144/000114238
ISSN0736-6205
Publisher versionhttp://www.biotechniques.com
Peer-Reviewedyes
AccessOpen access
Appears in Collections:CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series

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