dc.contributor.authorChan, Weng-Tat
dc.contributor.authorVerma, Chandra Shekhar
dc.contributor.authorLane, David P.
dc.contributor.authorGan, Samuel Ken-En
dc.date.accessioned2015-12-23T08:17:08Z
dc.date.available2015-12-23T08:17:08Z
dc.date.issued2013
dc.identifier.citationChan, W., Verma, C., Lane, D., & Gan, S. (2013). A comparison and optimization of methods and factors affecting the transformation of Escherichia coli. Bioscience Reports, 33(6), 931-937.en_US
dc.identifier.issn0144-8463en_US
dc.identifier.urihttp://hdl.handle.net/10220/39215
dc.description.abstractDNA manipulation routinely requires competent bacteria that can be made using one of numerous methods. To determine the best methods, we compared four commonly used chemical methods (DMSO, MgCl2–CaCl2, CaCl2 and Hanahan's methods) on frequently used Escherichia coli (E. coli) strains: DH5α, XL-1 Blue, SCS110, JM109, TOP10 and BL21-(DE3)-PLysS. Hanahan's method was found to be most effective for DH5α, XL-1 Blue and JM109 strains (P<0.05), whilst the CaCl2 method was best for SCS110, TOP10 and BL21 strains (P<0.05). The use of SOB (super optimal broth) over LB [Luria–Bertani (broth)] growth media was found to enhance the competency of XL-1 Blue (P<0.05), dampened JM109′s competency (P<0.05), and had no effect on the other strains (P>0.05). We found no significant differences between using 45 or 90 s heat shock across all the six strains (P>0.05). Through further optimization by means of concentrating the aliquots, we were able to get further increases in transformation efficiencies. Based on the optimized parameters and methods, these common laboratory E. coli strains attained high levels of TrE (transformation efficiency), thus facilitating the production of highly efficient and cost-effective competent bacteria.en_US
dc.description.sponsorshipASTAR (Agency for Sci., Tech. and Research, S’pore)en_US
dc.format.extent13 p.en_US
dc.language.isoenen_US
dc.relation.ispartofseriesBioscience Reportsen_US
dc.rights© 2013 The Author(s) This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC-BY) (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.en_US
dc.subjectCaCl2 methoden_US
dc.subjectCompetent bacteria
dc.subjectDMSO method
dc.subjectHanahan’s method
dc.subjectHeat shock
dc.titleA comparison and optimization of methods and factors affecting the transformation of Escherichia colien_US
dc.typeJournal Article
dc.contributor.schoolSchool of Biological Sciencesen_US
dc.identifier.doihttp://dx.doi.org/10.1042/BSR20130098
dc.description.versionPublished versionen_US


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record