chromosomes Protocols

Category: Cytogenetics Protocols

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

Category: Plant Biology Protocols: Plant Genetics Protocols

For analysis of metaphase chromosomes, any tissue containing dividing cells can be used: Root tips from young seedlings, from newly grown roots at the edge of plant pots or hydroponic culture are all suitable. Alternatively, flower buds, anthers, carpels or leaf or apical meristems can be used. Includes metaphase arresting reagents. - [Read Accumulation and Fixation of Plant Metaphase Chromosomes Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

The Allium test provides a rapid screening procedure for chemicals, pollutants contaminants, etc. which may represent environmental hazards.  Root growth inhibition and adverse effects upon chromosomes provide an indication of likely toxicity. - [Read Allium Test]

Category: Plant Biology Protocols

The Allium test provides a rapid screening procedure for chemicals, pollutants contaminants, etc. which may represent environmental hazards.  Root growth inhibition and adverse effects upon chromosomes provide an indication of likely toxicity. - [Read Allium Test]

Category: BACs Bacterial Artificial Chromosome Protocols

Information for BACs, Bacterial Artificial Chromosomes. Includes: Existing vectors; pBAC108L; pBeloBAC11; pBACe3.6; pFW11 and F' factor; pCC1BAC; pSMART VC Vectors; pIndigoBAC-5; Cosmids; pWEB-TNCTM; SuperCos 1; pFos1; PAC vectors; BAC purification; Putting the BAC into E. coli. - [Read BACs Bacterial Artificial Chromosomes]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Method is for preparing chromosomes from single flower buds of A. thaliana. It does not kill the plants allowing the determination of their chromosome number throughout development. Includes: Preparations of Arabidopsis; Preparation of Chromosome; Staining Chromosomes. - [Read Chromosome Spreads from Flower Buds of Arabidopsis thaliana Protocol]

Category: Cytogenetics Protocols

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)]

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Protocol describes a high sensitivity indirect detection procedure for DIG-labeled hybridization probes.  The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization.  This procedure can be used to detect single copy sequences as small as 1 kb on human metaphase chromosomes. 
    - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System Protocol]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

FISH Guide and Troubleshooting. Includes: Cytogenetic techniques; Slide preparation; When do chromosomes spread; Cell suspensions; G-banding; Cell fixatives; Slide preparation factors. - [Read FISH Guide and Troubleshooting]

Category: Plant Biology Protocols: Plant Genetics Protocols

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

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for fluorescence in situ hybridization (FISH) on human chromosomes and interphase nuclei. Includes: Plasmid probes; COSMID-, YAC- and LIBRARY PROBES; Probe detection. - [Read Fluorescence In Situ Hybridization (FISH) on Human Chromosomes and Interphase Nuclei Protocol]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for fluorescence in situ hybridization of a repetitive DNA probe to human chromosomes in suspension. Hybridization technique which does not need formamide and dextran sulfate. As a model system, we used the repetitive specific human DNA probe pUC 1.77, labeled it with digoxigenin-11-dUTP by nick-translation, and hybridized it to metaphase chromosomes in suspension. These chromosomes were isolated by standard techniques from human lymphocytes. - [Read Fluorescence In Situ Hybridization of a Repetitive DNA Probe to Human Chromosomes in Suspension]

Category: Fungi Protocols: Aspergillus Protocols

Compendium of protocols for using Aspergillus nidulans in genetic, molecular, and cell biological investigations, originally written for members of my research group. It also summarizes our common growth media and nutritional supplements, many of which originally appeared elsewhere but now are difficult to locate. Includes: Growth and storage of Aspergillus nidulans conidia; Nutritional supplements for our common auxotrophies; Double mutants; Mitotic mapping - assigning genes to chromosomes; etc - [Read Fundamentals of Growth, Storage, Genetics and Microscopy of Aspergillus nidulans Protocols]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to prepare aceto-orcein squashes, especially for pachytene chromosomes. - [Read How to Prepare Aceto-Orcein Squashes, Especially for Pachytene Chromosomes]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to separate chromosomes and restriction fragments using pulsed field gel electrophoresis. - [Read How to Separate Chromosomes and Restriction Fragments Using Pulsed Field Gel Electrophoresis]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol on how to stain Arabidopsis meiotic chromosomes from pollen meiocytes. - [Read How to Stain Arabidopsis Meiotic Chromosomes From Pollen Meiocytes Protocol]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to stain meiotic chromosomes using acriflavine. - [Read How to Stain Meiotic Chromosomes Using Acriflavine]

Category: Reporter Gene Assays: GFP Assay Protocols

Specific molecular components can be efficiently labeled by a combination of three methods: chemical transfection of GFP-fusion constructs, staining of chromosomes with the DNA-specific, fluorescent dye Hoechst 33342, and microinjection of fluorescently conjugated proteins. This procedure provides an example of using all three methods in sequence to label components of living HeLa cells. These methods should be followed in the order presented, but any of them can be omitted when not needed. - [Read Imaging Hoechst-Labeled Chromosomes and Fluorescent Proteins during the Cell Cycle]

Category: Microinjection Protocols

Fluorescence microscopy provides a powerful tool for imaging molecular components in living cells. Specific molecular components can be efficiently labeled by a combination of three methods: chemical transfection of GFP-fusion constructs, staining of chromosomes with the DNA-specific, fluorescent dye Hoechst 33342, and microinjection of fluorescently conjugated proteins. This procedure provides an example of using all three methods in sequence to label components of living HeLa cells. - [Read Imaging Hoechst-Labeled Chromosomes and Fluorescent Proteins during the Cell Cycle]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Protocol for In situ hybridization to human metaphase chromosomes using DIG-, biotin-, or fluorochrome-labeled DNA probes and detection with fluorochrome conjugates. Includes: Pretreatment of metaphase spreads on slides; Denaturation and hybridization; Single color fluorescent detection with immunological amplification; Multicolor fluorescence in situ hybridization (Multicolor FISH); Results obtained with human metaphase chromosome spreads. - [Read In Situ Hybridization to Human Metaphase Chromosomes using DIG-, Biotin- or Fluorochrome-Labeled DNA]

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Protocol for in situ hybridization to human metaphase chromosomes using DIG-, biotin-, or fluorochrome-labeled DNA probes and detection with fluorochrome conjugates. - [Read In Situ Hybridization to Human Metaphase Chromosomes Using DIG-, Biotin-, or Fluorochrome-Labeled]

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for integrating extrachromasomal arrays into the C. elegans chromosomes. - [Read Integrating Extrachromasomal Arrays into the C. elegans Chromosomes Protocol]

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Protocol for nonradioactive in situ hybridization to polytene chromosomes with a DIG-labeled DNA probe. Includes: Labeling the hybridization probe; Preparation and denaturation of polytene chromosomes from Drosophila, Chironomus, or other species; Hybridization and detection. - [Read Nonradioactive In Situ Hybridization to Polytene Chromosomes With a DIG-Labeled DNA Probe Protocol]

Category: Cytogenetics Protocols

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]

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol for preparation of DNA for pulsed-field gel electrophoresis: isolation of intact DNA from yeast. Yeast cells are first treated enzymatically to break down the cell walls and then resuspended in low-melting-temperature agarose plugs.  The DNA is liberated by infusing the plugs with lysis buffer and proteases.  This method is used to prepare both conventional and artificial yeast chromosomes. - [Read Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of Intact DNA from Yeast]