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dc.creatorMahmood,Tariq
dc.creatorZar,Tamkina
dc.creatorSaqlan Naqvi,S.M
dc.date2008-01-01
dc.date.accessioned2019-05-03T12:44:29Z
dc.date.available2019-05-03T12:44:29Z
dc.identifierhttps://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100015
dc.identifier.urihttp://revistaschilenas.uchile.cl/handle/2250/85000
dc.descriptionElectroporation entails brief, high intensity pulse to create transient pores in the cell membrane to facilitate the entry of exogenous macromolecules, which may otherwise be excluded. Removal of the external field leads to the resealing of the membrane electropores permitting the survival of the electrically stimulated recipient cells. Using this technique foreign deoxyribonucleic acid (DNA) has been successfully introduced into many cell types both from prokaryotes and eukaryotes. Increase in pulse voltage and length beyond a critical limit has been reported to decrease transformation efficiency, hence in this study we have investigated another strategy i.e. increase in the number of pulses at constant high voltage and pulse duration. Commonly used Agrobacterium strains LBA4404 and EHA101 and binary vector pCAMBIA1301 were used. Transformants were selected on a combination of hygromycin and kanamycin, and confirmed by polymerase chain reaction (PCR) and restriction analysis. Increase in the number of pulses was found to show a significant and linear increase in transformation efficiency.
dc.formattext/html
dc.languageen
dc.publisherPontificia Universidad Católica de Valparaíso
dc.relation10.4067/S0717-34582008000100015
dc.rightsinfo:eu-repo/semantics/openAccess
dc.sourceElectronic Journal of Biotechnology v.11 n.1 2008
dc.subjectAgrobacterium
dc.subjectelectroporation
dc.subjectpulse number
dc.subjecttransformation efficiency
dc.titleMultiple pulses improve electroporation efficiency in Agrobacterium tumefaciens


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