# Difference between revisions of "Cell cycle analysis"

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− | + | '''1.Determination of initiation age (a<sub>i</sub>) and C+D''':[[Image:Theoretical_age_distr.jpg|left|200px]] | |

From flow cytometry analysis of cells treated with rifampicin and cephalexin (run-out histogram) the proportions of cells that had not initiated replication at the time of drug action (4-origin-cells, streaked) and cells that had initiated (8-origin-cells) can be estimated.The initiation age (a<sub>i</sub>) can be found from the theoretical age distribution described by this formula, | From flow cytometry analysis of cells treated with rifampicin and cephalexin (run-out histogram) the proportions of cells that had not initiated replication at the time of drug action (4-origin-cells, streaked) and cells that had initiated (8-origin-cells) can be estimated.The initiation age (a<sub>i</sub>) can be found from the theoretical age distribution described by this formula, | ||

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[[Image:C+D_1.jpg|left|250px]] | [[Image:C+D_1.jpg|left|250px]] | ||

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+ | '''2. Determination of the C and D periods:''' | ||

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+ | The C period is found from the ''oriC/terC'' ratio obtained by Southern blot analysis and the generation time (τ): | ||

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+ | '''''oriC/terC''=2<sup>C/τ</sup>''' | ||

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+ | The D period is found from the C+D and C period: | ||

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+ | '''D = (C+D) - C''' | ||

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+ | Example (continues): | ||

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+ | C period calculated from the ''oriC/terC'' ratio: 49 min | ||

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+ | D period = (C+D) – C | ||

+ | D period = 76 min – 49 min = 27 min |

## Revision as of 07:31, 15 August 2008

**Cell cycle analysis of Escherichia coli cells**

C period = the time for a round of chromosome replication

D period = the time between the end of a round of chromosome replication and cell division

**1.Determination of initiation age (a**:

_{i}) and C+DFrom flow cytometry analysis of cells treated with rifampicin and cephalexin (run-out histogram) the proportions of cells that had not initiated replication at the time of drug action (4-origin-cells, streaked) and cells that had initiated (8-origin-cells) can be estimated.The initiation age (a_{i}) can be found from the theoretical age distribution described by this formula,

**F = 2 - 2 ^{((τ-ai)/τ)}**

where F is the fraction of cells that had not initiated and τ is the generation time, or from the estimated graph of the theoretical age distribution (streaked portion).

The C+D period is estimated from the initiation age (a_{i}), the generation time (τ) and the number of generations spanned per cell cycle.

Example:

4-origin-cells: 23 %

Generation time (τ): 27 min

Initiation age (a_{i}): 5 min

**2. Determination of the C and D periods:**

The C period is found from the *oriC/terC* ratio obtained by Southern blot analysis and the generation time (τ):

*oriC/terC*=2^{C/τ}

The D period is found from the C+D and C period:

**D = (C+D) - C**

Example (continues):

C period calculated from the *oriC/terC* ratio: 49 min

D period = (C+D) – C D period = 76 min – 49 min = 27 min