Show simple item record

dc.contributor.authorWood, Whitney N. ( )
dc.contributor.authorSmith, Kyle D. ( )
dc.contributor.authorReam, Jennifer A. ( )
dc.contributor.authorLewis, L. Kevin ( )
dc.date.accessioned2020-04-13T14:43:30Z
dc.date.available2020-04-13T14:43:30Z
dc.date.issued2017-02
dc.identifier.citationWood, W. N., Smith, K. D., Ream, J. A., & Lewis, L. K. (2017). Enhancing yields of low and single copy number plasmid DNAs from Escherichia coli cells. Journal of Microbiological Methods, 133, pp. 46–51.en_US
dc.identifier.issn0167-7012
dc.identifier.urihttps://digital.library.txstate.edu/handle/10877/9586
dc.description.abstractMany plasmids used for gene cloning and heterologous protein expression in Escherichia coli cells are low copy number or single copy number plasmids. The extraction of these types of plasmids from small bacterial cell cultures produces low DNA yields. In this study, we have quantitated yields of low copy and single copy number plasmid DNAs after growth of cells in four widely used broths (SB, SOC, TB, and 2xYT) and compared results to those obtained with LB, the most common E. coli cell growth medium. TB (terrific broth) consistently generated the greatest amount of plasmid DNA, in agreement with its ability to produce higher cell titers. The superiority of TB was primarily due to its high levels of yeast extract (24g/L) and was independent of glycerol, a unique component of this broth. Interestingly, simply preparing LB with similarly high levels of yeast extract (LB24 broth) resulted in plasmid yields that were equivalent to those of TB. By contrast, increasing ampicillin concentration to enhance plasmid retention did not improve plasmid DNA recovery. These experiments demonstrate that yields of low and single copy number plasmid DNAs from minipreps can be strongly enhanced using simple and inexpensive media.en_US
dc.formatText
dc.format.extent12 pages
dc.format.medium1 file (.pdf)
dc.language.isoen
dc.publisherElsevieren_US
dc.sourceJournal of Microbiological Methods, 2017, Vol. 133, pp. 46–51.
dc.subjectCopy numberen_US
dc.subjectDNA purification
dc.subjectLB
dc.subjectMiniprep
dc.subjectPlasmid
dc.subjectTB
dc.titleEnhancing Yields of Low and Single Copy Number Plasmid DNAs from E. coli Cellsen_US
txstate.documenttypeArticle
dc.description.versionThis is the accepted manuscript version of an article published in the Journal of Microbiological Methods.
dc.identifier.doihttps://doi.org/10.1016/j.mimet.2016.12.016
txstate.departmentChemistry and Biochemistry


Download

Thumbnail

This item appears in the following Collection(s)

Show simple item record