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Rapid, simple and potentially universal method for DNA extraction from Opuntia spp. fresh cladode tissues suitable for PCR amplification

2018Journal article Restricted access
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dc.contributor.authorRaimundo, Joana
dc.contributor.authorReis, C.M.G.
dc.contributor.authorRibeiro, M.M.A.
dc.date.accessioned2018-09-19T22:19:52Z
dc.date.available2018-09-19T22:19:52Z
dc.date.issued2018
dc.description.abstractIn Opuntia spp., the cladode tissues contain many polysaccharides and secondary metabolites that interfere with obtaining high-quality deoxyribonucleic acid (DNA), using currently available methods. To circumvent this problem, three commercial kits, three modified versions of the conventional cetyltrimethylammonium bromide method (CTAB) method and one combined method were tested in Opuntia ficus-indica, O. robusta, O. dillenii and O. elata species. We obtained a rapid and simple protocol that allows the extraction of DNA from all the tested species with good DNA yield and purity, namely, the combined method. With this method (DNeasy® Plant Mini Kit combined with the CTAB method), DNA yields from 13.2 ± 7.8 to 15.9 ± 11.3 µg.g-1 of fresh tissue were obtained in the four Opuntia species. The purity, evaluated by the ratio A260/A280 ratio, ranged from 1.67 ± 0.12 to 2.01 ± 0.25, revealing low levels of problematic metabolites. The extracted DNA quality was confirmed by amplifying a set of nuclear microsatellites obtained for the genus. Reliable reproducible bands and electropherogram profiles were obtained. The combined method has potential to be universal for good-quality DNA extraction in cacti, particularly in the Opuntia genus and other difficult-to-extract species.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationRaimundo J, Reis CMG, Ribeiro MM (2018) Rapid, simple and potentially universal method for DNA extraction from Opuntia spp. fresh cladode tissues suitable for PCR amplification. Molecular Biology Reports, in presspt_PT
dc.identifier.doi10.1007/s11033-018-4303-8pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.11/6172
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherSpringer Netherlandspt_PT
dc.relation.publisherversionhttps://link.springer.com/article/10.1007%2Fs11033-018-4303-8pt_PT
dc.subjectCactus pearpt_PT
dc.subjectCTABpt_PT
dc.subjectDNA extractionpt_PT
dc.subjectPolysaccharidespt_PT
dc.subjectViscositypt_PT
dc.titleRapid, simple and potentially universal method for DNA extraction from Opuntia spp. fresh cladode tissues suitable for PCR amplificationpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/UID%2FAGR%2F00239%2F2013/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/UID%2FAMB%2F00681%2F2013/PT
oaire.citation.titleMolecular Biology Reportspt_PT
oaire.fundingStream5876
oaire.fundingStream5876
person.familyNameRaimundo
person.familyNameGaspar Reis
person.familyNameRibeiro
person.givenNameJoana
person.givenNameCarlos
person.givenNameMaria Margarida
person.identifier349097
person.identifier.ciencia-idA71B-FF9D-83C6
person.identifier.ciencia-idAD12-4D32-7A48
person.identifier.orcid0000-0002-0850-4401
person.identifier.orcid0000-0002-4854-5128
person.identifier.orcid0000-0003-4684-1262
person.identifier.ridM-4235-2013
person.identifier.scopus-author-id7201715611
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsrestrictedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublicationa456aa66-5c6e-41f0-9501-38ff9e629ac7
relation.isAuthorOfPublication74d754d4-3926-497b-a0c4-26e98cbe0c93
relation.isAuthorOfPublicationf5b33ec3-9c90-4cc9-b0c4-c86c7ec0b017
relation.isAuthorOfPublication.latestForDiscovery74d754d4-3926-497b-a0c4-26e98cbe0c93
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