Cytogenetics Protocols

Karyotyping is a valuable research tool used to determine the chromosome complement within cultured cells. It is important to keep in mind that karyotypes evolve with continued culture. Because of this evolution, it is important for the interpretation of biochemical or other data, that the karyotype of a specific sub-line be determined. - [Read Karyotyping Protocol]

Protocol for staining chromosomes (G-Banding). Includes: Trypsin (1:250) USP Grade; Gurr Buffer Solution: (1 liter); Working Salt Solution: (1 liter); Leishman Stain Solution. - [Read Staining Chromosomes (G-Banding) Protocol]

Cytogenetic protocol for blood preps. Includes: Preparation of materials; Culture Procedure - Use aseptic technique throughout procedure until harvest; Harvest; Slide making; Collecting blood by tail vein method; PHA Dose Response. - [Read Cytogenetic Protocol Blood Preps]

Protocol for the preparation of metaphase spread. Includes three methods. - [Read Preparation of Metaphase Spread Protocol]

G-Banding observations. Includes: Slide storage; Fixative composition and trypsin treatment in G-banding; Banding of chemically aged slides. - [Read G-Banding Observations]

Protocol for accumulation and fixation of plant metaphase chromosomes. - [Read Accumulation and Fixation of Plant Metaphase Chromosomes Protocol]

MN in Human Lymphocytes (method and protocol description) Nina T. Holland, Division of Environmental Health Sciences, Berkeley - [Read Micronucleus assay Human Lymphocytes]

An ideal method of tissue preparation ensures both good specimen morphology and that the target molecules are in the optimum state for probe access and hybridization. DNA:DNA in situ hybridization is usually carried out on chromosome spread preparations where chromosome and nuclei are released from cells and spread on a glass microscope slide. This method yields well separated and enlarged chromosomes with good morphology which can be analyzed in transmitted light or fluorescence microscopes. - [Read Preparation of Chromosome Spreads]

Protocol for mitotic chromosome from cell cultures. Includes: Colchicine Treatment; Monolayer cultures; Cell Suspensions; Cell Harvest; Chromosome Slide Preparation. - [Read Mitotic Chromosome from Cell Cultures Protocol]

Protocol for Agrobacterium-mediated transformation in rice. Agrobacterium-mediated rice transformation method that is applicable to easily cultured varieties in addition to elite japonica varieties that are more difficult to culture. Using this method, transgenic rice plants can be obtained in about 2–3 months with a transformation frequency of 30–50%, both in easily cultured varieties and recalcitrant elite japonica rice. - [Read Agrobacterium-Mediated Transformation in Rice Protocol]

Protocol for mitotic chromosome preparations from bone marrow. - [Read Mitotic Chromosome Preparations from Bone Marrow Protocol]

Protocol for color changing karytoping (CCK): an M-FISH/SKY alternative. Includes: Slide preparation and hybridization; Probes, dyes and antibodies; Probe detection and image capture; Image processing. - [Read Color Changing Karyotyping (CCK) Protocol: An M-FISH/SKY Alternative]

Protocol for CGH of biotin labeled DNA vs digoxigenin labeled DNA. Includes: Reprecipitate DNA's; Denature slides; Hybridization; Washing and staining. - [Read CGH of Biotin Labeled DNA vs Digoxigenin Labeled DNA Protocol]

In situ hybridisation to alpha satellite sequences (chromosome specific). Method book .net - [Read In situ hybridisation to alpha satellite sequences (chromosome specific)]

Silver Staining for Nucleolar Organizer Regions. Trude Schwarzacher and Molecular Cytogenetics Research Group - [Read Silver Staining for Nucleolar Organizer Regions]

Protocol for mitotic chromosome preparations from mammalian suspension cell cultures. - [Read Mitotic Chromosome Preparations from Mammalian Suspension Cell Cultures Protocol]

CGH provides a comprehensive picture of all chromosomal gains and losses within a single experiment. In contrast to banding analysis, CGH requires no preparation of metaphase chromosomes from the cells to be analyzed (which in certain tumor types may be difficult or even impossible). - [Read Detection of Chromosomal Imbalances Using DOP-PCR and Comparative Genomic Hybridization (CGH)]

Protocol for pretreatment of chromosome slides for FISH/CGH/SKY. - [Read Pretreatment of Chromosome Slides for FISH/CGH/SKY Protocol]

Protocol describes a recently developed method — methylation-specific digital karyotyping (MSDK) — that enables comprehensive and unbiased genome-wide DNA methylation analysis. Using a combination of a methylation-sensitive mapping enzyme (for example, AscI) and a fragmenting enzyme (for example, NlaIII), short sequence tags can be obtained and uniquely mapped to genome location. - [Read Methylation-Specific Digital Karyotyping Protocol]

Protocol for meiotic preparations from Testis. - [Read Meiotic Preparations from Testis Protocol]

SCE protocol. Protocol for the use for wildtype MEFs. It may require some adjustments for other genotypes or cell types. - [Read SCE protocol]

Protocol for slide preparation for cytogenetics. Includes: Improved cell recovery by microcentrifugation; Slide storage; "Controlled" chromosome spreading; Dropping cells from a height does not improve spreading; GTG-banding. - [Read Slide Preparation for Cytogenetics Protocol]

Protocol for comparative genomic hybridization (CGH). Includes: Theory of CGH; Sensitivity; Applications of CGH. - [Read Comparative Genomic Hybridization (CGH) Protocol]

Protocol for nick translation using (CGH). - [Read Nick Translation (CGH) Protocol]

Protocol for DNA precipitation and hybridization (CGH). - [Read DNA Precipitation and Hybridization (CGH) Protocol]