chromosome Protocols

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

Describes an experimental cross in mice that can be used to define and map induced germ-line mutations that map to a single chromosome. The cross is a modification and extension of a conventional three-generation recessive mutagenesis screen. Includes: The Mutagenesis Breeding Plan; Consomic Strains; Generating Mutations; Generating and Genotyping G2 Females; Genotyping G3 Progeny; Phenotyping G4 Progeny; etc.. - [Read A Targeted Screen to Detect Recessive Mutations that have Quantitative Effects Protocol]

Category: BACs Bacterial Artificial Chromosome Protocols

Describe a set of BACs that map close to the centromeric and distal chromosomal ends for each mouse chromosome. - [Read A Triple Color FISH Technique for Mouse Chromosome Identification]

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol presents the amplification of insert end sequences from bacterial artificial chromosome clones using TAIL-PCR. The amplified products are suitable as probes for chromosome walking and genome mapping and as templates for direct sequencing. The protocol has been used in rice genome studies. - [Read Amplification of Insert End Sequences from BACs Clones by Thermal Asymmetric Interlaced PCR]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for chromosome preparation and Fluorescence in-situ hybridization (FISH). Chromosome preparation and Fluorescence in-situ hybridization (FISH). - [Read Chromosome Preparation and Fluorescence In-Situ Hybridization (FISH) Protocol]

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: Genetics and Genomics: Cancer Genetics Protocols

Investigators can utilize X chromosome inactivation (methylation) to determine the clonality status of a tumor or premalignant lesion in females. The technique is based on a methylation-sensitive restriction enzyme and analysis of a polymorphic locus on the X chromosome. Clonal cell populations will show "loss" of the non-methylated allele after restriction digest. The assay can be performed on DNA recovered from microdissected samples. Both frozen tissue and fixed-embedded tissue can be used. - [Read Clonality - X Chromosome Inactivation Assay Protocol]

Category: Real Time PCR

Genotyping Ts65Dn mice is based on doing simultaneous quantitative PCR amplification of a gene or genes in the Ts65Dn chromosome and a control gene on another chromosome (in this case Apob) and comparing the average change (delta) in threshold cycle (CT) - [Read Genotyping Mice Using Real Time]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to obtain and recognize partial-diploid strains that are duplicated for known chromosome segments. - [Read How to Obtain and Recognize Partial-Diploid Strains that are Duplicated for Known Chromosome Segment]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to recognize and diagnose chromosome rearrangements. - [Read How to Recognize and Diagnose Chromosome Rearrangements]

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast PCR Protocols

To identify the YAC subclones containing both a human insert and a portion of either the left or right arm of the pYAC4 vector. Identification of these clones is necessary in order to do YAC chromosome walking, and is also useful in the determination of whether a particular YAC clone has a contiguous human insert or whether a co-cloning event has occurred. Vector arm sequences are identified using pBR322 fragments from a BamHI-PvuII double digest. - [Read Identification of End Clones in YAC Subclone Libraries Protocol]

Category: Cytogenetics Protocols

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

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

In situ hybridisation to alpha satellite sequences (chromosome specific) protocol. - [Read In Situ Hybridisation to Alpha Satellite Sequences (Chromosome Specific) Protocol]

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

Protocol for in situ hybridisation to alpha satellite sequences (chromosome specific). - [Read In Situ Hybridisation to Alpha Satellite Sequences Protocol]

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: 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]

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol describes the use of agarose plugs for isolation of yeast artificial chromosome (YAC) DNA. The DNA can then be run on a pulsed-field gel and used for microinjection to produce transgenic mice. - [Read Large-Scale Preparation of Agarose Plugs of Yeast DNA Protocol]

Category: Cytogenetics Protocols

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]

Category: Cytogenetics Protocols

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

Category: Cytogenetics Protocols

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

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol for the preparation of BAC (Bacterial Artificial Chromosome) DNA with CONCERT high purity plasmid purification system. Includes: - [Read Preparation of BAC DNA with CONCERT High Purity Plasmid Purification System 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: Cytogenetics Protocols

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

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

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

Category: Cytogenetics Protocols: Spectral Karyotyping SKY Protocols

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

Category: Fungi Protocols: Neurospora Protocols

Protocol guide for the N. crassa yeast artificial chromosome library. Includes: Chromosome Walking; Hybridization screening of the YAC library; YAC restriction mapping and contig building; Preparation of chromosomal DNA plugs of YAC clones; Partial restriction enzyme digestion of YAC DNA plugs; Using CHEF gel analysis to resolve YAC clones; Southern Hybridization; Isolation of terminal restriction fragments from cloned DNA inserts in YAC clones; etc. - [Read Protocol Guide for the N. crassa Yeast Artificial Chromosome Library]