Cell Cycle Analysis by DNA Content
  • OVERVIEW

Cell Cycle Analysis by DNA Content


Cell cycle analysis by DNA content measurement is a method that most frequently employs flow cytometry to distinguish cells in different phases of the cell cycle. Before analysis, the cells are usually permeabilised and treated with a fluorescent dye that stains DNA quantitatively, such as propidium iodide (PI) or 4,6-diamidino-2-phenylindole (DAPI). The fluorescence intensity of the stained cells correlates with the amount of DNA they contain. As the DNA content doubles during the S phase, the DNA content (and thereby intensity of fluorescence) of cells in the G0 phase and G1 phase (before S), in the S phase, and in the G2 phase and M phase (after S) identifies the cell cycle phase position in the major phases (G0/G1 versus S versus G2/M phase) of the cell cycle. The cellular DNA content of individual cells is often plotted as their frequency histogram to provide information about relative frequency (percentage) of cells in the major phases of the cell cycle.

Propidium iodide (PI) is a fluorescent dye that binds to DNA. When excited by 488nm laser light, it can be detected with in the PE/Texas Red® channel with a bandpass filter 610/10. It is commonly used in evaluation of cell viability or DNA content in cell cycle analysis by flow cytometry.


Workflow of Our Service

-Receiving orders and samples

-Transfection of target vectors

-Cell collection

-Fixation

-Removal of fixative

-Staining

-Flow cytometry analysis

-Project report writing and delivery

Sample Requirements

Cell line: >1x10^6 cells.

Please use dry ice to transport the sample.

Deliverables

Raw data, figures, analysis results and project report including our methods and materials.

Cell Cycle Analysis by DNA Content


Cell cycle analysis by DNA content measurement is a method that most frequently employs flow cytometry to distinguish cells in different phases of the cell cycle. Before analysis, the cells are usually permeabilised and treated with a fluorescent dye that stains DNA quantitatively, such as propidium iodide (PI) or 4,6-diamidino-2-phenylindole (DAPI). The fluorescence intensity of the stained cells correlates with the amount of DNA they contain. As the DNA content doubles during the S phase, the DNA content (and thereby intensity of fluorescence) of cells in the G0 phase and G1 phase (before S), in the S phase, and in the G2 phase and M phase (after S) identifies the cell cycle phase position in the major phases (G0/G1 versus S versus G2/M phase) of the cell cycle. The cellular DNA content of individual cells is often plotted as their frequency histogram to provide information about relative frequency (percentage) of cells in the major phases of the cell cycle.

Propidium iodide (PI) is a fluorescent dye that binds to DNA. When excited by 488nm laser light, it can be detected with in the PE/Texas Red® channel with a bandpass filter 610/10. It is commonly used in evaluation of cell viability or DNA content in cell cycle analysis by flow cytometry.


Workflow of Our Service

-Receiving orders and samples

-Transfection of target vectors

-Cell collection

-Fixation

-Removal of fixative

-Staining

-Flow cytometry analysis

-Project report writing and delivery

Sample Requirements

Cell line: >1x10^6 cells.

Please use dry ice to transport the sample.

Deliverables

Raw data, figures, analysis results and project report including our methods and materials.

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