| Z701866 Sigma |
Cell Cycle Checkpoint Control Protocols |
General description
A collection of cutting-edge techniques for studying the mechanisms underlying cell cycle regulation and checkpoint control. Using mammalian, yeast, and frog systems, these readily reproducible methods can be used to induce cell cycle checkpoints, detect changes in cell cycle progression, identify and analyze genes and proteins that regulate the process, and characterize chromosomal status as a function of cell cycle phase and progression. Each fully tested technique includes step-by-step instructions, an introduction explaining the principle behind the method, equipment and reagent lists, and tips on troubleshooting and avoiding known pitfalls
Table of Contents
PART I. Induction and Detection of Changes in Cell Cycle Progression
1. Methods to Induce Cell Cycle Checkpoints
2. Methods for Synchronizing Mammalian Cells
3. Enrichment of Cells in Different Phases of the Cell Cycle by Centrifugal Elutriation
4. Analysis of the Mammalin Cell Cycle by Flow Cytometry
5. Methods for Detecting Cell in S Phase
6. Yeast Cell Synchronization
7. Analysis of the Budding Yeast Saccharomyces cerevisiae Cell Cycle by Morphological Criteria and Flow Cytometry
8. Analysis of the Fission Yeast Schizosaccharomyces pombe Cell Cycle
PART II. Analysis of Genes Involved in Checkpoint Control
9. Strategies to Isolate Evolutionaarily Conserved Cell Cycle Regulatory Genes
10. Microarray Approaches for Analysis of Cell Cycle Regulatory Genes
11. Using the Yeast Genome-Wide Gene-Deletion Collection for Systematic Genetic Screens
12. Gene Targeting in Cultured Human Cells
13. Use of In Vivo Gap Repair for Isolation of Mutatn Alleles of a Checkpoint Gene
14. In Vitro Mutagenesis to Define Functional Domains
15. Use of Gene Overexpression to Assess Function in Cell Cycle Control
16. Histone Acetylation/Deacetylation As a Regulator of Cell Cycle Gene Expression
PART III. Analysis of Proteins Involved in Checkpoint Control
17. Cataloging Proteins in Cell Cycle Control
18. Multidimentsional Proteomic Analysis of Proteolytic Pathways Involved in Cell Cycle Control
19. Purification and Identification of Protein Complexes That Control the Cell Cycle
20. Xenopus Cell-Free Extracts to Study DNA Damage Checkpoints
21. Protein-Protein Interactions
22. Detection of Kinase and Phosphatase Activities
23. Monitoring Changes in the Subcellular Location of Proteins in S. cerevisiae
PART IV. Chromosomes and the Cell Cycle
24. Chromosomal Changes and the Cell Cycle Checkpoints in Mammalian Cells
25. Detecting the Influence of Cell Cycle Regulatory Proteins on Human Telomeres
26. Monitoring spindle Assembly and Disassembly in Yeast by Indirect Immunofluorescence
Index
1. Methods to Induce Cell Cycle Checkpoints
2. Methods for Synchronizing Mammalian Cells
3. Enrichment of Cells in Different Phases of the Cell Cycle by Centrifugal Elutriation
4. Analysis of the Mammalin Cell Cycle by Flow Cytometry
5. Methods for Detecting Cell in S Phase
6. Yeast Cell Synchronization
7. Analysis of the Budding Yeast Saccharomyces cerevisiae Cell Cycle by Morphological Criteria and Flow Cytometry
8. Analysis of the Fission Yeast Schizosaccharomyces pombe Cell Cycle
PART II. Analysis of Genes Involved in Checkpoint Control
9. Strategies to Isolate Evolutionaarily Conserved Cell Cycle Regulatory Genes
10. Microarray Approaches for Analysis of Cell Cycle Regulatory Genes
11. Using the Yeast Genome-Wide Gene-Deletion Collection for Systematic Genetic Screens
12. Gene Targeting in Cultured Human Cells
13. Use of In Vivo Gap Repair for Isolation of Mutatn Alleles of a Checkpoint Gene
14. In Vitro Mutagenesis to Define Functional Domains
15. Use of Gene Overexpression to Assess Function in Cell Cycle Control
16. Histone Acetylation/Deacetylation As a Regulator of Cell Cycle Gene Expression
PART III. Analysis of Proteins Involved in Checkpoint Control
17. Cataloging Proteins in Cell Cycle Control
18. Multidimentsional Proteomic Analysis of Proteolytic Pathways Involved in Cell Cycle Control
19. Purification and Identification of Protein Complexes That Control the Cell Cycle
20. Xenopus Cell-Free Extracts to Study DNA Damage Checkpoints
21. Protein-Protein Interactions
22. Detection of Kinase and Phosphatase Activities
23. Monitoring Changes in the Subcellular Location of Proteins in S. cerevisiae
PART IV. Chromosomes and the Cell Cycle
24. Chromosomal Changes and the Cell Cycle Checkpoints in Mammalian Cells
25. Detecting the Influence of Cell Cycle Regulatory Proteins on Human Telomeres
26. Monitoring spindle Assembly and Disassembly in Yeast by Indirect Immunofluorescence
Index
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