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A ten-year search for synchronous cells: obstacles, solutions, and practical applications.

Helmstetter CE - Front Microbiol (2015)

Bottom Line: My effort to use synchronously dividing cultures to examine the Escherichia coli cell cycle involved a 10-year struggle with failure after failure punctuated by a few gratifying successes, especially at the end.In this essay, I recount my personal journey in this obsessive experimental pursuit.That narrative is followed by a description of a simplified version of the "baby machine," a technique that was developed to obtain minimally disturbed, synchronously growing E. coli cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Florida Institute of Technology Melbourne, FL, USA.

ABSTRACT
My effort to use synchronously dividing cultures to examine the Escherichia coli cell cycle involved a 10-year struggle with failure after failure punctuated by a few gratifying successes, especially at the end. In this essay, I recount my personal journey in this obsessive experimental pursuit. That narrative is followed by a description of a simplified version of the "baby machine," a technique that was developed to obtain minimally disturbed, synchronously growing E. coli cells. Subsequent studies with this methodology led to an understanding of the basic properties of the relationship between chromosome replication and cell division. Accordingly, I end this reminiscence with a simple, fool-proof graphical strategy for deducing the pattern of chromosome replication during the division cycle of cells growing at any rate.

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Genome equivalents of DNA per cell (G) during the division cycle. Calculations of the values for G are shown in red for three select times in the division cycle: 0, 10, and 15 min.
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Figure 7: Genome equivalents of DNA per cell (G) during the division cycle. Calculations of the values for G are shown in red for three select times in the division cycle: 0, 10, and 15 min.

Mentions: Finally, it is sometimes of interest to determine chromosomal DNA content per cell at various times in the cycle in terms of genome equivalents (G) (Cooper and Helmstetter, 1968). This calculation is shown in red in Figure 7 for three select times in the division cycle (0, 10, and 15 min). Again based on the idea that the replication rate is constant during C, this calculation is most easily visualized by recording the extent of chromosomal replication based on time rather than distance, and then dividing the sum of the times by C (40 min in this case) as shown. The average chromosomal DNA content in an exponentially growing culture can be determined with the equation: , where τ equals doubling time.


A ten-year search for synchronous cells: obstacles, solutions, and practical applications.

Helmstetter CE - Front Microbiol (2015)

Genome equivalents of DNA per cell (G) during the division cycle. Calculations of the values for G are shown in red for three select times in the division cycle: 0, 10, and 15 min.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4373376&req=5

Figure 7: Genome equivalents of DNA per cell (G) during the division cycle. Calculations of the values for G are shown in red for three select times in the division cycle: 0, 10, and 15 min.
Mentions: Finally, it is sometimes of interest to determine chromosomal DNA content per cell at various times in the cycle in terms of genome equivalents (G) (Cooper and Helmstetter, 1968). This calculation is shown in red in Figure 7 for three select times in the division cycle (0, 10, and 15 min). Again based on the idea that the replication rate is constant during C, this calculation is most easily visualized by recording the extent of chromosomal replication based on time rather than distance, and then dividing the sum of the times by C (40 min in this case) as shown. The average chromosomal DNA content in an exponentially growing culture can be determined with the equation: , where τ equals doubling time.

Bottom Line: My effort to use synchronously dividing cultures to examine the Escherichia coli cell cycle involved a 10-year struggle with failure after failure punctuated by a few gratifying successes, especially at the end.In this essay, I recount my personal journey in this obsessive experimental pursuit.That narrative is followed by a description of a simplified version of the "baby machine," a technique that was developed to obtain minimally disturbed, synchronously growing E. coli cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Florida Institute of Technology Melbourne, FL, USA.

ABSTRACT
My effort to use synchronously dividing cultures to examine the Escherichia coli cell cycle involved a 10-year struggle with failure after failure punctuated by a few gratifying successes, especially at the end. In this essay, I recount my personal journey in this obsessive experimental pursuit. That narrative is followed by a description of a simplified version of the "baby machine," a technique that was developed to obtain minimally disturbed, synchronously growing E. coli cells. Subsequent studies with this methodology led to an understanding of the basic properties of the relationship between chromosome replication and cell division. Accordingly, I end this reminiscence with a simple, fool-proof graphical strategy for deducing the pattern of chromosome replication during the division cycle of cells growing at any rate.

No MeSH data available.


Related in: MedlinePlus