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Validation of reference genes for the normalization of RT-qPCR gene expression in Acanthamoeba spp.

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cris.virtual.author-orcid#PLACEHOLDER_PARENT_METADATA_VALUE#
cris.virtual.author-orcid#PLACEHOLDER_PARENT_METADATA_VALUE#
cris.virtual.author-orcid#PLACEHOLDER_PARENT_METADATA_VALUE#
cris.virtual.author-orcid#PLACEHOLDER_PARENT_METADATA_VALUE#
cris.virtualsource.author-orcid#PLACEHOLDER_PARENT_METADATA_VALUE#
cris.virtualsource.author-orcid#PLACEHOLDER_PARENT_METADATA_VALUE#
cris.virtualsource.author-orciddf1c65d5-8f4e-48b2-a70a-2bc13c2e09b2
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dc.contributor.authorKöhsler, Martina
dc.contributor.authorLeitsch, David
dc.contributor.authorMüller, Norbert
dc.contributor.authorWalochnik, Julia
dc.date.accessioned2024-09-02T17:09:02Z
dc.date.available2024-09-02T17:09:02Z
dc.date.issued2020-06-25
dc.description.abstractAcanthamoebae are potentially pathogenic organisms, with a highly unique, yet still insufficiently investigated metabolism. Many open questions can be addressed by gene expression studies, however, for Acanthamoeba reliable standards have not yet been established. In this study, suitable reference genes (RGs) for RT-qPCR in Acanthamoeba were comprehensively evaluated, comparing different Acanthamoeba strains and employing four different algorithms (NormFinder, GeNorm, BestKeeper and RefFinder). Expression stability was assessed under various conditions and the potentials of the most promising RGs for accurate normalization of target genes were evaluated. Expression stability of RGs varied depending on conditions and employed algorithms, however, the genes for the 18S rRNA and the hypoxanthine phosphoribosyl transferase seem to be widely suitable RGs. Normalization with a combination of two carefully chosen RGs resulted in reliable expression data for target genes, while normalization with unsuitable RGs led to significant misinterpretation of expression profiles. Thus, a careful evaluation of RGs prior to expression studies is essential.
dc.description.sponsorshipInstitut für Infektionskrankheiten, Parasitologie
dc.identifier.doi10.48350/154117
dc.identifier.pmid32587282
dc.identifier.publisherDOI10.1038/s41598-020-67035-0
dc.identifier.urihttps://boris-portal.unibe.ch/handle/20.500.12422/40992
dc.language.isoen
dc.publisherNature Publishing Group
dc.relation.ispartofScientific Reports
dc.relation.issn2045-2322
dc.relation.organizationDCD5A442BA1AE17DE0405C82790C4DE2
dc.relation.organizationDCD5A442BD12E17DE0405C82790C4DE2
dc.relation.organizationDCD5A442BFE6E17DE0405C82790C4DE2
dc.relation.organizationDCD5A442C1CCE17DE0405C82790C4DE2
dc.subject.ddc500 - Science::570 - Life sciences; biology
dc.subject.ddc600 - Technology::610 - Medicine & health
dc.subject.ddc600 - Technology::630 - Agriculture
dc.titleValidation of reference genes for the normalization of RT-qPCR gene expression in Acanthamoeba spp.
dc.typearticle
dspace.entity.typePublication
oaire.citation.issue1
oaire.citation.startPage10362
oaire.citation.volume10
oairecerif.author.affiliation#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliationInstitut für Infektionskrankheiten, Parasitologie
oairecerif.author.affiliation#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation2#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation2#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation2#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation2#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation3#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation3#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation3#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation3#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation4#PLACEHOLDER_PARENT_METADATA_VALUE#
oairecerif.author.affiliation4#PLACEHOLDER_PARENT_METADATA_VALUE#
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oairecerif.author.affiliation4#PLACEHOLDER_PARENT_METADATA_VALUE#
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unibe.contributor.rolecreator
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unibe.date.licenseChanged2021-03-17 15:11:47
unibe.description.ispublishedpub
unibe.eprints.legacyId154117
unibe.journal.abbrevTitleSci Rep
unibe.refereedTRUE
unibe.subtype.articlejournal

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