Immobilization of microalgae cells in alginate facilitates isolation of DNA and RNA

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dc.contributor.authorLopez, Blanca R.
dc.contributor.authorHernández Sánchez, Juan Pablo
dc.contributor.authorBashan, Yoav
dc.contributor.authorde-Bashan, Luz E.
dc.contributor.orcidHernández Sánchez, Juan Pablo [0000-0003-1175-0109]
dc.date.accessioned2020-08-03T14:28:33Z
dc.date.available2020-08-03T14:28:33Z
dc.date.issued2017
dc.description.abstractenglishIsolation of nucleic acids from Chlorella is difficult, given the chemically complex nature of their cell walls and variable production of metabolites. Immobilization of microalgae in polymers adds additional difficulty. Here, we modified, amended, and standardized methods for isolation of nucleic acids and compared the yield of DNA and RNA from free-living and encapsulated microalgae C. sorokiniana. Isolation of nucleic acids from immobilized cells required two steps in dissolving the alginate matrix, releasing the cells, and mechanical disruption with glass beads. For DNA extraction, we used modified versions of a commercial kit along with the hexadecyltrimethylammonium bromide (CTAB) method. For RNA extraction, we used the commercial TRI reagent procedure and the CTAB-dithiotreitol method. Quantity and quality of nucleic acids in extracts varied with growth conditions, isolation procedures, and time of incubation of the original culture. There were consistently higher amounts of DNA and RNA in extracts from immobilized cells. Quantitatively, the modified procedure with the commercial Promega kit was the most reliable procedure for isolating DNA and a modified commercial TRI reagent procedure was the choice for isolating RNA. All four procedures eliminated proteins efficiently and had low levels of contamination from residual polysaccharides from the matrices and/or metabolites naturally produced by the microalgae. All DNA extracts under both growth conditions, time of incubation, and two isolation methods successfully amplified the 18S ribosomal RNA by PCR and quantitative reverse transcription (RT-qPCR).eng
dc.format.mimetypeapplication/pdf
dc.identifier.doihttps://doi.org/10.1016/j.mimet.2017.02.005
dc.identifier.instnameinstname:Universidad El Bosquespa
dc.identifier.issn0167-7012
dc.identifier.reponamereponame:Repositorio Institucional Universidad El Bosquespa
dc.identifier.repourlhttps://repositorio.unbosque.edu.co
dc.identifier.urihttps://hdl.handle.net/20.500.12495/3649
dc.language.isoeng
dc.publisherElsevierspa
dc.publisher.journalJournal of Microbiological Methodsspa
dc.relation.ispartofseriesJournal of Microbiological Methods, 0167-7012, Vol 135, 2017, pag 96-104spa
dc.relation.ispartofseriesJournal of Microbiological Methods, 0167-7012, Vol. 135, 2017, p. 96-104spa
dc.relation.urihttps://www.sciencedirect.com/science/article/pii/S0167701217300453?via%3Dihub
dc.rights.accessrightshttps://purl.org/coar/access_right/c_abf2
dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.rights.accessrightsAcceso abierto
dc.rights.creativecommons2017-04
dc.rights.localAcceso abiertospa
dc.subject.keywordsChlorellaspa
dc.subject.keywordsImmobilizationspa
dc.subject.keywordsMicroalgaespa
dc.subject.keywordsNucleic acidspa
dc.titleImmobilization of microalgae cells in alginate facilitates isolation of DNA and RNAspa
dc.title.translatedImmobilization of microalgae cells in alginate facilitates isolation of DNA and RNAspa
dc.type.coarhttps://purl.org/coar/resource_type/c_6501
dc.type.driverinfo:eu-repo/semantics/article
dc.type.hasversioninfo:eu-repo/semantics/publishedVersion
dc.type.localArtículo de revista

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