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Mitosis and meiosis. Part A /
| Clasificación: | Libro Electrónico |
|---|---|
| Otros Autores: | , |
| Formato: | Electrónico eBook |
| Idioma: | Inglés |
| Publicado: |
Camridge, MA :
Academic Press,
2018.
|
| Colección: | Methods in cell biology ;
v. 144. |
| Temas: | |
| Acceso en línea: | Texto completo Texto completo |
Tabla de Contenidos:
- Front Cover; Mitosis and Meiosis Part A; Copyright; Contents; Contributors; Preface; Chapter 1: Assays for the spindle assembly checkpoint in cell culture; 1. Introduction; 2. Choice of Fluorescent Protein Tag; 3. Choice of Microscope; 4. Assays; 5. Materials; Acknowledgments; References; Chapter 2: Quantitative methods to measure aneuploidy and chromosomal instability; 1. Introduction; 2. Immunofluorescence Assay for Lagging Chromosomes in Anaphase; 2.1. Instrumentation; 2.2. Cell Culture Conditions; 2.3. Antibodies; 2.4. Immunofluorescence; 2.5. Analysis; 2.6. Alternative Method
- 3. Chromosome Missegregation Assay3.1. Instrumentation; 3.2. Cell Culture Conditions; 3.3. FISH Probes; 3.4. Preparation of Mitotic Cells; 3.5. FISH Procedure; 3.6. Analysis; 3.7. Alternative Method; 4. Aneuploid Cell Survival Assay; 4.1. Instrumentation; 4.2. Cell Culture Conditions; 4.3. FISH Probes; 4.4. Preparation of Interphase Cells; 4.5. FISH Procedure; 4.6. Analysis; 4.7. Alternative Method; 5. Concluding Remarks; Acknowledgments; References; Chapter 3: Dissecting the role of the tubulin code in mitosis; 1. Introduction; 1.1. What Is the Tubulin Code?; 1.2. Tubulin Isotypes
- 1.3. Tubulin PTMs1.4. How Is the Tubulin Code Read?; 2. Modulation of the Detyrosination/Tyrosination Cycle in Mammalian Cells; 2.1. Cell Culture; 2.2. Transient Overexpression of TTL; 2.3. Depletion of TTL Using Small Interference RNAs (siRNAs); 2.4. Knockout of TTL Using CRISPR/Cas9; 2.4.1. Purchase oligos; 2.4.2. Oligo annealing and cloning into viral transfer vectors; 2.4.3. Transformation and selection; 2.4.4. Lentivirus production; 2.4.5. Transduction of lentivirus to target cells; 2.4.6. Selection of knockout cells; 2.5. Transient Overexpression of VASH1 and VASH2
- 2.6. Generation of Cell Lines Stably Expressing FLAG-VASH1 and FLAG-VASH22.6.1. Retrovirus production; 2.6.2. Transduction of retrovirus to target cells and selection of overexpressing cells; 2.7. Knockout of VASH1 and VASH2 Using CRISPR/Cas9; 2.7.1. Purchase oligos; 2.7.2. Oligo annealing and cloning into viral transfer vectors; 2.7.3. Lentivirus production; 2.7.4. Transduction of lentivirus to target cells and selection of knockout cells; 2.8. Depletion of Endogenous �I�-Tubulin Isotypes Using siRNA; 2.9. Transient Overexpression of Tyrosinated, Detyrosinated and Delta2 Forms of TUBA1B
- 2.9.1. Site-directed mutagenesis of mammalian expression vectors2.9.2. Altering the cDNA sequences to confer resistance to siRNA depletion; 2.9.3. Transient expression of tyrosinated, detyrosinated, and Delta2 forms of TUBA1B; 2.10. Generation of a Cell Line Stably Expressing H2B-mRFP and Tyrosinated, Detyrosinated, or Delta2 Forms of TUBA1B; 2.10.1. Cloning of TUBA1B cDNA into lentiviral transfer vectors; 2.10.2. Deletion of EGFP-tag from lentiviral vectors expressing TUBA1B by PCR; 2.10.3. Production of lentivirus and transduction to target cells