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pELMO, an optimised in-house cloning vector

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dc.creatorRamos, Andrea E.spa
dc.creatorMuñoz, Marinaspa
dc.creatorMoreno‑Pérez, Darwin A.spa
dc.creatorPatarroyo, Manuel A.spa
dc.creator.googleRamos, Andrea E.spa
dc.creator.googleMuñoz, Marinaspa
dc.creator.googleMoreno‑Pérez, Darwin A.spa
dc.creator.googlePatarroyo, Manuel A.spa
dc.date.accessioned2018-12-14T13:02:55Z
dc.date.available2018-12-14T13:02:55Z
dc.date.created2017spa
dc.date.issued2017
dc.description.abstractDNA cloning is an essential tool regarding DNA recombinant technology as it allows the replication of foreign DNA fragments within a cell. pELMO was here constructed as an in-house cloning vector for rapid and low-cost PCR product propagation; it is an optimally designed vector containing the ccdB killer gene from the pDONR 221 plasmid, cloned into the pUC18 vector’s multiple cloning site (Thermo Scientific). The ccdB killer gene has a cleavage site (CCC/GGG) for the SmaI restriction enzyme which is used for vector linearisation and cloning blunt-ended products. pELMO transformation efficiency was evaluated with different sized inserts and its cloning efficiency was compared to that of the pGEM-T Easy commercial vector. The highest pELMO transformation efficiency was observed for ~500 bp DNA fragments; pELMO vector had higher cloning efficiency for all insert sizes tested. In-house and commercial vector cloned insert reads after sequencing were similar thus highlighting that sequencing primers were designed and localised appropriately. pELMO is thus proposed as a practical alternative for in-house cloning of PCR products in molecular biology laboratories. © 2017, The Author(s).eng
dc.format.mimetypeapplication/pdf
dc.identifier.issn21910855
dc.identifier.urihttp://repository.urosario.edu.co/handle/10336/18824
dc.language.isoengspa
dc.relation.citationIssueNo. 1
dc.relation.citationTitleAMB Express
dc.relation.citationVolumeVol. 7
dc.relation.ispartofAMB Express, ISSN: 2191-0855, Vol. 7/No. 1 (2017)spa
dc.relation.urihttps://amb-express.springeropen.com/track/pdf/10.1186/s13568-017-0324-2spa
dc.rights.accesRightsinfo:eu-repo/semantics/openAccess
dc.rights.accesoAbierto (Texto completo)spa
dc.rights.cchttp://creativecommons.org/licenses/by/4.0/spa
dc.source.bibliographicCitationAinsa, J.A., Martin, C., Cabeza, M., De la Cruz, F., Mendiola, M.V., Construction of a family of Mycobacterium/Escherichia coli shuttle vectors derived from pAL5000 and pACYC184: their use for cloning an antibiotic-resistance gene from Mycobacterium fortuitum (1996) Gene, 176 (1-2), pp. 23-26. , COI: 1:CAS:528:DyaK28XntVKrtbo%3D, PID: 8918226spa
dc.source.instnameinstname:Universidad del Rosario
dc.source.reponamereponame:Repositorio Institucional EdocUR
dc.subjectBlunt-Endedspa
dc.subjectCcdb Killer Genespa
dc.subjectCloning Vectorspa
dc.subjectPcr Cloningspa
dc.subjectRecombinant Dna Technologyspa
dc.subject.decsPlasmid Vectorspa
dc.subject.decsRestriction Endonucleasespa
dc.subject.decsArticlespa
dc.subject.decsBacterium Transformationspa
dc.subject.decsCloning Vectorspa
dc.subject.decsMolecular Cloningspa
dc.subject.decsNeospora Caninumspa
dc.subject.decsNonhumanspa
dc.subject.decsPelmo Vectorspa
dc.subject.decsPlasmodium Falciparumspa
dc.subject.decsPlasmodium Vivaxspa
dc.subject.decsPolymerase Chain Reactionspa
dc.subject.decsProcess Designspa
dc.subject.decsProcess Optimizationspa
dc.subject.decsProtein Cleavagespa
dc.subject.decsSanger Sequencingspa
dc.titlepELMO, an optimised in-house cloning vectorspa
dc.typearticleeng
dc.type.hasVersioninfo:eu-repo/semantics/publishedVersion
dc.type.spaArtículospa
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