Chromosome Analysis Protocols by John R. Gosden

By John R. Gosden

Transparent, step by step directions give you all of the concepts utilized in either classical and molecular cytogenetics. The emphasis is at the use of human fabric, however the recommendations are designed to be without difficulty appropriate to different animal chromosomes. Chapters characteristic equipment utilized in the coaching of chromosomes, in studying chromosomes without delay on the light-microscope point, and in molecular genetics strategies.

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Left: a metaphase plate. Scale bar =lO pm. Right: a single chromosome from the same metaphase. Scale bar = 1 pm. been possible to combine in situ hybridization with high-quality morphological preservation. As well as providing alternative approaches to chromosome morphology, the methods using cytocentrifuge preparation and isolated chromosomes have the advantage of using little or no fixation prior to the osmium impregnation procedure. They are, therefore, well suited to the study of chromosomal antigens that might be destroyed or extracted by methanol-acetic acid fixation (Fig.

Ethidium Bromtde Technique (Originally Described by Ikeuchi [2U. 1. When mamtammg cells for making chromosomes by this method, the cells are kept m a semiconfluent state and only split when the flask is fully confluent (stationary phase). This will give a very crude but reasonably effective means of synchronizing rapidly growing cells. However, the chromosomes are in a more nearly “native” state after this. 2. Seventeen to 20 h after splitting the flask, add 100 JJL of ethtdium bromide and 100 pL of colcemid.

Chromosoma 100,4 10-4 18. 4. Harrison, C. J , Jack, E M , and Allen, T. , (1987) Light and scanningelectron microscopy of the same metaphase chromosomes, m Correlative Microscopy in Biology. ) Academic, New York, pp. 189-248. 5 Harrison, C J , Jack, E. M , Allen, T. , and Harris, R. (1983) The fragile X* a scanning electron microscope study. , and Harris, R (1985) Investtgation of human chromosome polymorphisms by scanning electron mtcroscopy. J. Med Genet 22,16-23 7. Sumner, A. T. (1986) Electron microscopy of the parameres formed by the centromeric heterochromatin of human chromosome 9 at pachytene.

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