Näytä suppeat kuvailutiedot

dc.contributor.authorMäki, Anita
dc.contributor.authorTiirola, Marja
dc.date.accessioned2018-10-17T05:51:21Z
dc.date.available2018-10-17T05:51:21Z
dc.date.issued2018
dc.identifier.citationMäki, A., & Tiirola, M. (2018). Directional high-throughput sequencing of RNAs without gene-specific primers. <i>BioTechniques</i>, <i>65</i>(4), 219-223. <a href="https://doi.org/10.2144/btn-2018-0082" target="_blank">https://doi.org/10.2144/btn-2018-0082</a>
dc.identifier.otherCONVID_28653850
dc.identifier.otherTUTKAID_79091
dc.identifier.urihttps://jyx.jyu.fi/handle/123456789/59844
dc.description.abstractRibosomal RNA analysis is a useful tool for characterization of microbial communities. However, the lack of broad-range primers has hampered the simultaneous analysis of eukaryotic and prokaryotic members by amplicon sequencing. We present a complete workfl ow for directional, primer-independent sequencing of size-selected small subunit ribosomal RNA fragments. The library preparation protocol includes gel extraction of the target RNA, ligation of an RNA oligo to the 5′-end of the target, and cDNA synthesis with a tailed random-hexamer primer and further barcoding. The sequencing results of a phytoplankton mock community showed a highly similar profi le to the biomass indicators. This method has universal potential for microbiome studies, and is compatible for the 5′-end sequencing of other RNA types with minimum library preparation costs.fi
dc.format.mimetypeapplication/pdf
dc.language.isoeng
dc.publisherFuture Science
dc.relation.ispartofseriesBioTechniques
dc.rightsCC BY-NC-ND 4.0
dc.subject.otherligation
dc.subject.othernext-generation sequencing
dc.subject.otherphytoplankton
dc.subject.otherprimer bias
dc.subject.otherrandom priming
dc.subject.otherribosomal RNA
dc.titleDirectional high-throughput sequencing of RNAs without gene-specific primers
dc.typearticle
dc.identifier.urnURN:NBN:fi:jyu-201810084365
dc.contributor.laitosBio- ja ympäristötieteiden laitosfi
dc.contributor.laitosDepartment of Biological and Environmental Scienceen
dc.contributor.oppiaineYmpäristötiedefi
dc.contributor.oppiaineNanoscience Centerfi
dc.contributor.oppiaineEnvironmental Scienceen
dc.contributor.oppiaineNanoscience Centeren
dc.type.urihttp://purl.org/eprint/type/JournalArticle
dc.date.updated2018-10-08T06:15:12Z
dc.type.coarhttp://purl.org/coar/resource_type/c_2df8fbb1
dc.description.reviewstatuspeerReviewed
dc.format.pagerange219-223
dc.relation.issn0736-6205
dc.relation.numberinseries4
dc.relation.volume65
dc.type.versionpublishedVersion
dc.rights.copyright© Authors, 2018
dc.rights.accesslevelopenAccessfi
dc.relation.grantnumber615146
dc.relation.grantnumber615146
dc.relation.projectidinfo:eu-repo/grantAgreement/EC/FP7/615146/EU//
dc.subject.ysomikrobisto
dc.subject.ysoRNA
dc.subject.ysoribosomit
dc.subject.ysolevät
dc.subject.ysogeenitekniikka
dc.subject.ysomikrolevät
dc.subject.ysoplankton
dc.format.contentfulltext
jyx.subject.urihttp://www.yso.fi/onto/yso/p27039
jyx.subject.urihttp://www.yso.fi/onto/yso/p7689
jyx.subject.urihttp://www.yso.fi/onto/yso/p22179
jyx.subject.urihttp://www.yso.fi/onto/yso/p3327
jyx.subject.urihttp://www.yso.fi/onto/yso/p17994
jyx.subject.urihttp://www.yso.fi/onto/yso/p26977
jyx.subject.urihttp://www.yso.fi/onto/yso/p3053
dc.rights.urlhttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.relation.doi10.2144/btn-2018-0082
dc.relation.funderEuroopan komissiofi
dc.relation.funderEuropean Commissionen
jyx.fundingprogramEU:n 7. puiteohjelma (FP7)fi
jyx.fundingprogramFP7 (EU's 7th Framework Programme)en
jyx.fundinginformationAM was supported by the European Research Council (ERC) through a grant awarded to MT under the European Union’s Seventh Framework Programme (FP/2007-2013, grant 615146). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.
dc.type.okmA1


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