Aneuploidy:
The chromosomal constitution of cells which deviate from the normal by the addition or subtraction of Chromosomes, chromosome pairs, or chromosome fragments. In a normally diploid cell (Diploidy) the loss of a chromosome pair is termed nullisomy (symbol: 2N-2), the loss of a single chromosome is Monosomy (symbol: 2N-1), the addition of a chromosome pair is tetrasomy (symbol: 2N+2), the addition of a single chromosome is Trisomy (symbol: 2N+1).
JoVE General
Chromosomics: Detection of Numerical and Structural Alterations in All 24 Human Chromosomes Simultaneously Using a Novel OctoChrome FISH Assay
Genes and Environment Laboratory, University of California, Berkeley
A novel fluorescence in situ hybridization (FISH) method that simultaneously examines both numerical and structural chromosome alterations, particularly the specific chromosomal translocations associated with leukemia and lymphoma, of all 24 human chromosomes on a single device in one hybridization, is described.
JoVE General
Fluorescence in situ hybridization (FISH) Protocol in Human Sperm
Unitat de Biologia Cel·lular (Facultat de Biociències), Universitat Autónoma de Barcelona
This video-article describes, step by step, how to process a semen sample to achieve good-quality fluorescence in situ hybridization on human spermatozoa. Preparations obtained can be used for aneuploidy screening in the context of clinical diagnosis.
JoVE General
Mouse Oocyte Microinjection, Maturation and Ploidy Assessment
Department of Biology, University of Pennsylvania
Oocytes are prone to aneuploidy due to errors in chromosome segregation during meiotic maturation. Aneuploid eggs can cause infertility, miscarriages or developmental disorders like Down syndrome. Here, we describe methods to introduce materials of choice into oocytes and methods to study meiotic maturation and assess ploidy.
JoVE General
Studying Mitotic Checkpoint by Illustrating Dynamic Kinetochore Protein Behavior and Chromosome Motion in Living Drosophila Syncytial Embryos
Institute for Cell and Molecular Biosciences, University of Newcastle, United Kingdom
The kinetochore is where the SAC initiates its signal monitoring the mitotic segregation of the sister chromatids. A method is described to visualize the recruitment and turnover of one of the kinetochore proteins and its coordination with the chromosome motion in Drosophila embryos using a Leica laser scanning confocal system.
JoVE Clinical and Translational Medicine
FISH for Pre-implantation Genetic Diagnosis
This article describes the selection of suitable probes for single-cell FISH, spreading techniques for blastomere nuclei, and in situ hybridization and signal scoring, applied to pre-implantation genetic diagnosis (PGD) in a clinical setting.
JoVE General
Chromosomal Spread Preparation of Human Embryonic Stem Cells for Karyotyping
Institute of Biomedical Sciences, Federal University of Rio De Janeiro-UFRJ
Karyotyping is a simple and useful technique widely used for detecting genetic alterations. Here we describe a step by step protocol for chromosome spread preparation of human embryonic stem cells for monitoring the chromosomal status of these cells maintained in culture.
JoVE General
MISSION esiRNA for RNAi Screening in Mammalian Cells
Max Planck Institute of Molecular Cell Biology and Genetics
Here we use a human esiRNA library in a high-throughput screen for genes involved in cell division. We demonstrate how to set up and conduct an esiRNA screens, as well as how to analyze and validate the results.
JoVE General
The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions
1Department of Synthetic Biology and Bioenergy, J. Craig Venter Institute, 2Department of Microbial and Environmental Genomics, J. Craig Venter Institute, 3Donnelly Centre & Department of Molecular Genetics, University of Toronto, 4Lunenfeld Research Institute, Mt Sinai Hospital
The Green Monster method enables the rapid assembly of multiple deletions marked with a reporter gene encoding green fluorescent protein. This method is based on driving yeast strains through repeated cycles of sexual assortment of deletions and fluorescence-based enrichment of cells carrying more deletions.
JoVE General
Studying Proteolysis of Cyclin B at the Single Cell Level in Whole Cell Populations
Department of Hematology, Oncology and Stem Cell Transplantation, University Medical Center Freiburg
Metaphase to anaphase transition is triggered through anaphase-promoting complex (APC/C)-dependent ubiquitination and subsequent destruction of cyclin B. Here, we established a system which, following pulse-chase labeling, allows monitoring cyclin B proteolysis in entire cell populations and facilitates the detection of interference by the mitotic checkpoint.
JoVE General
Design and Use of Multiplexed Chemostat Arrays
Department of Genome Sciences, University of Washington
We developed and validated a small-footprint array of miniature chemostats built from readily available parts for low cost. Physiological and experimental evolution results were similar to larger volume chemostats. The ministat array provides a compact, inexpensive, and accessible platform for traditional chemostat experiments, functional genomics, and chemical screening applications.
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