Rapid determination of plasmid copy number

Schmidt T, Friehs K, Flaschel E (1996)

Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
Plasmid copy number, the number of expression vectors per host cell, is a key variable in recombinant microbial cultivation. Therefore, it would be very helpful, if the plasmid copy number could be determined during the operating process period. A rapid quantification of this important process variable would even open the possibility of its use in process control. However, current assays like gel electrophoresis, CsCl-gradient centrifugation, HPLC and other methods are time consuming and difficult to quantify. Indirect methods: like the correlation of copy number with, e.g. the activity of an enzyme, coded on the plasmid, are prone to errors due to the production kinetics, turnover rate and protein denaturation. Here, a method is presented, which enables the plasmid copy number to be determined in less than 30 min. This novel procedure based on plasmid isolation by means of a commercial DNA-isolation kit and quantification by capillary electrophoresis, should allow the copy number to be used in process control.
DNA quantification; plasmid copy number; plasmid isolation; electrophoresis; recombinant Escherichia; capillary; coli; laser induced fluorescence
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Schmidt T, Friehs K, Flaschel E. Rapid determination of plasmid copy number. JOURNAL OF BIOTECHNOLOGY. 1996;49(1-3):219-229.
Schmidt, T., Friehs, K., & Flaschel, E. (1996). Rapid determination of plasmid copy number. JOURNAL OF BIOTECHNOLOGY, 49(1-3), 219-229. https://doi.org/10.1016/0168-1656(96)01519-2
Schmidt, T, Friehs, Karl, and Flaschel, Erwin. 1996. “Rapid determination of plasmid copy number”. JOURNAL OF BIOTECHNOLOGY 49 (1-3): 219-229.
Schmidt, T., Friehs, K., and Flaschel, E. (1996). Rapid determination of plasmid copy number. JOURNAL OF BIOTECHNOLOGY 49, 219-229.
Schmidt, T., Friehs, K., & Flaschel, E., 1996. Rapid determination of plasmid copy number. JOURNAL OF BIOTECHNOLOGY, 49(1-3), p 219-229.
T. Schmidt, K. Friehs, and E. Flaschel, “Rapid determination of plasmid copy number”, JOURNAL OF BIOTECHNOLOGY, vol. 49, 1996, pp. 219-229.
Schmidt, T., Friehs, K., Flaschel, E.: Rapid determination of plasmid copy number. JOURNAL OF BIOTECHNOLOGY. 49, 219-229 (1996).
Schmidt, T, Friehs, Karl, and Flaschel, Erwin. “Rapid determination of plasmid copy number”. JOURNAL OF BIOTECHNOLOGY 49.1-3 (1996): 219-229.

14 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Quantification of Plasmid Copy Number with Single Colour Droplet Digital PCR.
Plotka M, Wozniak M, Kaczorowski T., PLoS One 12(1), 2017
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Capacitive biosensor for quantification of trace amounts of DNA.
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Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes.
Skulj M, Okrslar V, Jalen S, Jevsevar S, Slanc P, Strukelj B, Menart V., Microb Cell Fact 7(), 2008
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The copy-number of plasmids and other genetic elements can be determined by SYBR-Green-based quantitative real-time PCR.
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PMID: 16216354
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