Chromosome Painting: Principles, Strategies and Scope by Arun Kumar Sharma, Archana Sharma

By Arun Kumar Sharma, Archana Sharma

Chromosome portray is the main sleek and novel method for without delay settling on numerous gene sequences at the same time within the chromosome, through particular probes in molecular hybridization. Its answer levels from unmarried replica to whole genome sequences. it's now utilized in plant, animal, and human platforms, in gene mapping, id of genetic problems, evolutionary reports, and gene move experiments. This treatise is the 1st of its sort to hide the method with all its transformations and functions. it truly is designed for normal use by way of postgraduate scholars and examine staff in mobile and molecular genetics, plant and animal sciences, agriculture, drugs, and phylogenetic studies.

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Chromosome Painting: Principles, Strategies and Scope

Chromosome portray is the main smooth and novel strategy for without delay choosing numerous gene sequences at the same time within the chromosome, through particular probes in molecular hybridization. Its answer levels from unmarried reproduction to whole genome sequences. it's now utilized in plant, animal, and human platforms, in gene mapping, id of genetic problems, evolutionary reports, and gene move experiments.

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Probe preparation and labelling 01. Extract DNA from yeast and bacteria according to standard methods [12]. 02 . If YAC clone s are used, amplify the human inserts by inter-Alu PCR as described by Hoglund et al. [8], using the primers ILA 3' and 5' . 03 . Label I ug of extracted DNA/PCR product by incorporation of hapten- or fluorochromecoupled nucleotides using the Megaprime kit. Label each probe in a separate tube. Denature the DNA together with 5 III primer solution in boiling water for 5 min.

Cremer T . Murken J. Spei cher MR (1999). Mu ltiplex-FISH for pre- and pos tna tal diag nosti c appl icat ions. Am J Hum Genet 65: 448-462. 14. Verma RS . Babu A (199 5) . Hu man Chromosomes : princi ples and tech niq ues . New York: McG raw -Hil I. Address for Corre spondence: David Gisselsson . Depart ment of Clinical Genetics . University Hospital. SE · 22 1 85 Lun d. se Methods in Cell Science 23: 29-35 (2001) . © 2001 Kluwer Academic Publishers. Printed in the Netherlands. Relationship of telomere sequence and constitutive heterochromatin in the human and apes as detected by PRINS Hirohisa Hirai Primate Research Institute, Kyoto Univer sity, Center for Human Evolutionary Modeling Research and Department of Cellular and Molecular Biology, Inuyama, Aichi 484 -8506, Japan Abstract.

Centrifuge at 14,000 rpm for 20 min and remo ve the supernatant. Dissolve the labelled DNA in 50% form amide/l 0% de xtran sulfate/2x SSC to a concentration of 20-50 ng/ul and store at -20 °C until used. C. Chromosome preparation for FISH 01. Culture the cell s in a 25 ern? plastic flask in a suitable medium. 02. 5-3 h at 37 °C. 03. Pour all the culture medium into a labelled 10 ml test tube . 085 M trisodium citrate solution to the culture fla sk and let stand for I min at RT. 04. 8 % trypsin to the flask.

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