Chromosome Analysis ProtocolsJohn R. Gosden Springer Science & Business Media, 2 feb 2008 - 509 páginas Chromosomes, as the genetic vehicles, provide the basic material for a large proportion of genetic investigations, from the construction of gene maps and models of chromosome organization, to the inves tigation of gene function and dysfunction. The study of chromosomes has developed in parallel with other aspects of molecular genetics, beginning with the first preparations of chromosomes from animal cells, through the development of banding techniques, which permitted the unequivocal identification of each chromosome in a karyotype, to the present analytical methods of molecular cytogenetics. Although some of these techniques have been in use for many years, and can be learned relatively easily, most published scientific reports—as a result of pressure on space from editors, and the response to that pressure by authors—contain little in the way of technical detail, and thus are rarely adequate for a researcher hoping to find all the necessary information to embark on a method from scratch. A new user needs not only a detailed description of the methods, but also some help with problem solving and sorting out the difficulties en countered in handling any biological system. This was the require ment to which the series Methods in Molecular Biology is addressed, and Chromosome Analysis Protocols forms a part of this series. |
Índice
11 | |
Meiotic Chromosome Preparation | 27 |
Preparation of Chromosomes for Scanning Electron Microscopy | 41 |
Chromosome Preparation and Banding | 51 |
Chromosome Banding and Identification Absorption Staining | 59 |
Fluorescence | 83 |
Stain Combinations for Specific Regions | 97 |
Inhibition of Chromosome Condensation | 113 |
vii | 231 |
Immunofluorescence Techniques Applied to Mitotic Chromosome | 253 |
Immunocytochemical Techniques Applied to Meiotic Chromosomes | 287 |
Their Creation and Use in Gene Mapping | 303 |
Electron Microscopic Localization of in Situ Hybrids | 335 |
Construction of ChromosomeSpecific Libraries of Yeast Artificial | 379 |
Yeast Artificial Chromosome Recombinants in a Global Strategy | 403 |
Chromosome Dissection and Cloning | 425 |
Analysis of Chromosomes with Restriction Endonucleases | 123 |
Automatic Karyotype Analysis | 141 |
Bivariate Chromosome Analysis Using a Commercial Flow | 187 |
Production of | 205 |
Pulsed Field Gel Electrophoresis | 221 |
Molecular Analysis of Chromosome Aberrations | 437 |
Alu and L1Primed PCRGenerated Probes for Nonisotopic | 479 |
Telomeres | 493 |
505 | |
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Términos y frases comunes
2X SSC agarose gel aliquot amplification antibody antigens buffer cell lines centrifuge centromere Chapter chro chromo Chromomycin Chromosome Analysis chromosome banding chromosome preparations clones CMGT colcemid concentration coverslip culture Cytogenet DAPI denatured described detection digestion diluted distilled water EDTA electrophoresis ethanol ethidium bromide filter FITC flow karyotypes fluorescence fluorochromes fragments gene Genet genome Giemsa heterochromatin Hoechst human chromosomes hybridization immunofluorescence Incubate karyotyping labeled mapping markers medium meiotic membrane metaphase metaphase chromosomes method mg/mL microscope mitotic Molecular mosome mouse NaCl Natl nick translation Note pellet pipet pixels primers probe procedure prophase Protocols R-banding reaction restriction enzyme Resuspend Rinse room temperature sample Section sequences serum Sigma signal situ hybridization slides sodium sorbitol spheroplasts Spin staining sterile stock solution store at 20°C supernatant suspension technique telomeres tion transfer Tris-HCl tube vector Wash yeast artificial chromosome µg/mL