Cell Cycle Analysis Protocols

Information on propidium iodide for cell cycle analysis. - [Read Cell Cycle Analysis - Propidium Iodide]

This protocol describes a method for the synchronization of cell populations using centrifugal elutriation. The method relies on the fact that cell size increases linearly as cells proceed through the cell cycle. Cells of similar size (and cell cycle phase) are eluted stepwise from the cell chamber, with the smallest size (those in early G1) being eluted first. Using this procedure, it is possible to obtain relatively pure populations of cells at various points in G1, S, and G2/M. - [Read Cell Synchronization Using Centrifugal Elutriation Protocol]

Describes assays used to determine the distribution of a population of cells to the different stages of the cell cycle as analyzed by flow cytometry. Staining the DNA with different fluorescent dyes, propidium iodide or DAPI, is one of the most direct ways of staging the cells based on DNA content. - [Read Determining Cell Cycle Stages by Flow Cytometry Protocol]

This protocol provides a method for the synchronization of a monolayer culture of CHO cells in G1 using isoleucine deprivation. Since CHO cells can also be adapted to grow in suspension culture, this procedure can be used to obtain larger quantities of cells. When isoleucine is replaced, the cells resume growth and begin to enter S phase ~4 hours later. This method arrests almost 100% of the CHO cells in G1, and upon reversal, leads to rapid recovery of cell growth and very high cell viability. - [Read G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol]

This protocol provides a method for synchronizing cells at the G1/S border using a double treatment of thymidine, which, in excess, is an inhibitor of DNA synthesis. Cells are treated once with excess thymidine to accumulate the majority of them at G1/S; however, some cells will have stopped growth within the S phase. - [Read G1/S Phase Synchronization using Double Thymidine Synchronization Protocol]

This protocol uses the plant amino acid mimosine as a G1/S synchronizing agent. Cells are first treated with excess thymidine to accumulate the majority of them at G1/S; however, some cells will have stopped growth within the S phase. Thymidine is then removed to allow all the cells to proceed completely through the S phase. Mimosine is then added to arrest the cells at the G1/S border. When mimosine is removed, cells will begin to enter S phase within about 1 hour. - [Read G1/S Phase Synchronization Using Mimosine Arrest Protocol]

Protocol for H-Thymidine uptake by cultured cells. - [Read H-Thymidine Uptake by Cultured Cells Protocol]

Exponentially growing cells are asynchronous with respect to the cell cycle stage. Detection of cell cycle-related events is improved by enriching the culture for cells at the stage during which the particular event occurs. Methods for synchronizing cells are provided here, including those based on morphological features of the cell. - [Read Methods for Synchronizing Cells at Specific Stages of the Cell Cycle]

Protocol on mitosis. - [Read Mitosis Protocol]

Protocol for preparing cells for PI/FACS (cell cycle) analysis. - [Read Preparing Cells for PI/FACS (cell cycle) Analysis Protocol]

Protocol describes methods for maintaining healthy, dividing mammalian cells in culture and during imaging, when mitosis can be examined. Rose chambers are preferable for observation and microinjection of living mitotic cells, but slide/coverslip preparations are easy to make and do not require any special equipment. - [Read Studying Mitosis in Cultured Mammalian Cells Protocol]

This protocol describes a method for synchronizing monolayer cells in mitosis using selective detachment from their substrate. During mitosis, cells become more spherical, causing them to become more loosely attached to their substrate. The "rounded up" cells are selectively detached by tapping the culture flask, resulting in a population in which as many as 90-98% of the cells are in mitosis. The drug nocodazole is used to increase the percentage of cells undergoing mitosis before detachment.. - [Read Synchronization of Mammalian Cell Cultures in Mitosis Using Selective Detachment Protocol]

Protocol for timing of cycles. - [Read Timing of Cycles Protocol]

Protocol for yeast cell cycle by flow cytometry. - [Read Yeast Cell Cycle by Flow Cytometry Protocol]