Repository logo
 
Publication

Plasmid production and purification: An integrated experiment‐based biochemistry and biotechnology laboratory course

2019Journal article Restricted access
Simple item page
dc.contributor.authorSantos, Tiago
dc.contributor.authorPereira, Patrícia
dc.contributor.authorQueiroz, João
dc.contributor.authorCruz, Carla
dc.contributor.authorSousa, Fani
dc.date.accessioned2020-01-09T17:23:31Z
dc.date.available2020-01-09T17:23:31Z
dc.date.issued2019
dc.description.abstractThis laboratory experiment describes the production and purification of plasmid DNA for undergraduate biochemistry and biotechnology courses. This experiment performed in a one-week period includes the protocols for plasmid pVAX1-LacZ production in Escherichia coli DH5α cells and subsequent purification of supercoiled pVAX1-LacZ. Firstly, the students use a growth medium that favors the replication of the plasmid resulting in a higher plasmid production during exponential growth. Afterwards, alkaline lysis is done to disrupt the bacterial cells and recover pVAX1-LacZ plasmid. Lastly, they perform the purification of pVAX1-LacZ supercoiled isoform by L-histidine chromatography, followed by agarose gel electrophoresis to characterize the separation of supercoiled isoform from contaminants. The proposed experiment provides an opportunity for students to acquire these skills that are routinely used in biochemistry and biotechnology laboratories. © 2019 International Union of Biochemistry and Molecular Biology, 47(6):638-643, 2019.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1002/bmb.21290pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/8186
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherWileypt_PT
dc.relationDRug dElivery nAnosysteM for HPV infection therapy
dc.relation.publisherversionhttps://iubmb.onlinelibrary.wiley.com/doi/full/10.1002/bmb.21290pt_PT
dc.subjectBiochemistrypt_PT
dc.subjectBiotechnologypt_PT
dc.subjectDNA - Bacterialpt_PT
dc.subjectEscherichia colipt_PT
dc.subjectHumanspt_PT
dc.subjectLaboratoriespt_PT
dc.subjectPlasmidspt_PT
dc.subjectStudentspt_PT
dc.subjectUniversitiespt_PT
dc.subjectCurriculumpt_PT
dc.titlePlasmid production and purification: An integrated experiment‐based biochemistry and biotechnology laboratory coursept_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleDRug dElivery nAnosysteM for HPV infection therapy
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5665-PICT/MIT-EXPL%2FBIO%2F0008%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5665-PICT/UTAP-EXPL%2FNTec%2F0015%2F2017/PT
oaire.citation.endPage643pt_PT
oaire.citation.issue6pt_PT
oaire.citation.startPage638pt_PT
oaire.citation.titleBiochemistry and Molecular Biology Educationpt_PT
oaire.citation.volume47pt_PT
oaire.fundingStream5665-PICT
oaire.fundingStream5665-PICT
person.familyNameAfonso dos Santos
person.familyNamePereira
person.familyNameQueiroz
person.familyNameCruz
person.familyNameSousa
person.givenNameTiago André
person.givenNamePatrícia
person.givenNameJoão
person.givenNameCarla
person.givenNameFani
person.identifier.ciencia-id6517-B7D0-2B07
person.identifier.ciencia-id4715-067B-4FD0
person.identifier.ciencia-id931E-B66D-E341
person.identifier.ciencia-id6816-7888-F8B1
person.identifier.ciencia-id991D-2E13-A840
person.identifier.orcid0000-0003-4620-0174
person.identifier.orcid0000-0002-5665-5271
person.identifier.orcid0000-0002-3096-8325
person.identifier.orcid0000-0001-6630-1242
person.identifier.orcid0000-0001-9996-2194
person.identifier.ridL-3104-2014
person.identifier.ridI-7806-2013
person.identifier.ridA-2014-2017
person.identifier.scopus-author-id56111844900
person.identifier.scopus-author-id7003705645
person.identifier.scopus-author-id7202701394
person.identifier.scopus-author-id7005110268
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.embargofctCopyright cedido à editora no momento da publicaçãopt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublicationb21d9803-f183-4785-b060-49c7859b4c17
relation.isAuthorOfPublication157c4d6b-e0e0-4ef2-9d25-e461156ced0c
relation.isAuthorOfPublicationc798c033-c160-4f99-a8ad-04416923943f
relation.isAuthorOfPublication3fc51bf5-f849-4292-9aa3-4947fac1b9c8
relation.isAuthorOfPublication2935de97-fc90-4d38-9505-4caa364f8a10
relation.isAuthorOfPublication.latestForDiscovery2935de97-fc90-4d38-9505-4caa364f8a10
relation.isProjectOfPublicationf48e17e4-0dcc-41b3-9e93-f8753ccb30bc
relation.isProjectOfPublicationbc2bd40f-d998-44de-a1c1-42760dcd6bc8
relation.isProjectOfPublication.latestForDiscoveryf48e17e4-0dcc-41b3-9e93-f8753ccb30bc

Files

Original bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
Santos et al_Biochem Mol Biol Educ_2019.pdf
Size:
1.25 MB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.71 KB
Format:
Item-specific license agreed upon to submission
Description:
Download

Collections

ICI - CICS | Documentos por Auto-Depósito
FCS - DCM | Documentos por Auto-Depósito