Use of the Sucrose Gradient Method for Bacterial Cell Cycle Synchronization
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Authors:
Lin Lin1,
Abha Choudhary1,
Anish Bavishi1,
Norma Ogbonna1,
Sarah Maddux1,
Madhusudan Choudhary1,*
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Published 03 May 2012
- *Corresponding author. Mailing address: Department of Biological Sciences, Sam Houston State University, Avenue I, Lee Drain Building, Suite 300, P.O. Box 2116, Huntsville, Texas 77341. Phone: 936-294-4850. Fax: 936-294-3940. E-mail: mchoudhary@shsu.edu.
- Copyright © 2012 American Society for Microbiology
Abstract:
Although many undergraduate and graduate Cell and Molecular Biology courses study the bacterial cell cycle and the mechanisms that regulate prokaryotic cell division, few laboratory projects exist for the enhanced study of cell cycle characteristics in a standard teaching laboratory. One notable reason for this lack of engaging laboratory projects is, although bacterial cells can be grown fairly easily, these cultured cells are in a variety of cell cycle states. As such, to study and understand the factors that regulate bacterial cell division in morphological, physiological, and even molecular respects, it is necessary to have bacterial cells in the same stage of its cell cycle. This matching can be performed by a procedure called cell cycle synchronization.
Key Concept Ranking
- Scanning Electron Microscopy
- 0.46891212
- Bacterial Cell Division
- 0.4550528
References & Citations
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Abstract:
Although many undergraduate and graduate Cell and Molecular Biology courses study the bacterial cell cycle and the mechanisms that regulate prokaryotic cell division, few laboratory projects exist for the enhanced study of cell cycle characteristics in a standard teaching laboratory. One notable reason for this lack of engaging laboratory projects is, although bacterial cells can be grown fairly easily, these cultured cells are in a variety of cell cycle states. As such, to study and understand the factors that regulate bacterial cell division in morphological, physiological, and even molecular respects, it is necessary to have bacterial cells in the same stage of its cell cycle. This matching can be performed by a procedure called cell cycle synchronization.

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Author and Article Information
-
Published 03 May 2012
- *Corresponding author. Mailing address: Department of Biological Sciences, Sam Houston State University, Avenue I, Lee Drain Building, Suite 300, P.O. Box 2116, Huntsville, Texas 77341. Phone: 936-294-4850. Fax: 936-294-3940. E-mail: mchoudhary@shsu.edu.
- Copyright © 2012 American Society for Microbiology
Figures
Buoyant densities of R. sphaeroides under different growth conditions. The determination of buoyant densities of R. sphaeroides cells both aerobically (left panel) and photosynthetically (right panel).

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FIGURE 1
Buoyant densities of R. sphaeroides under different growth conditions. The determination of buoyant densities of R. sphaeroides cells both aerobically (left panel) and photosynthetically (right panel).
R. sphaeroides cells separated into two different sucrose gradient layers with corresponding buoyant densities.

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FIGURE 2
R. sphaeroides cells separated into two different sucrose gradient layers with corresponding buoyant densities.
Microscopic analysis of cells collected from different gradient layers and synchronized pellets: (A) cells with majority of dividing cells were collected from 68% density layers; (B) cells with evenly mixed cell populations were taken from 69% layers; (C) cells were collected from bottom pellets with highest buoyant densities. Note: Most cells were newborn.

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FIGURE 3
Microscopic analysis of cells collected from different gradient layers and synchronized pellets: (A) cells with majority of dividing cells were collected from 68% density layers; (B) cells with evenly mixed cell populations were taken from 69% layers; (C) cells were collected from bottom pellets with highest buoyant densities. Note: Most cells were newborn.