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FISH+slides+2011 - Karyotyping Which chromosomes are there...

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1 Karyotyping Which chromosomes are there? Are they normal? Tools of the trade Gene identification by breakpoint Changes in chromosome structure and number by FISH Fluorescence In Situ Hybridization Old style banding allows identification of each chromosome by bands and size http://www.sciencemuseum.org.uk/exhibitions/genes/154.asp Chromosome painting and banding allows identification of each chromosome Specific color pattern for each chromosome One color per chromosome http://www.bio.unc.edu/courses/2003spring/biol050h/biolog2.jpg Spectral karyotyping (SKY) is a laboratory technique that allows scientists to visualize all of the human chromosomes at one time by "painting" each pair of chromosomes in a different fluorescent color. www.genome.gov/10000208 Interphase vs metaphase DOTS vs BANDS 3 sep probes INTERPHASE IS CHEAPER AND EASIER vs colchicine - arrested metaphases Interphase vs metaphase probes on same chromosome are close 2 probes on same chromosome are close to each other
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2 Detecting aberrations Abberations and FISH www.rerf.or.jp/ Gene/eng/fig/fishf.htm Chromosome painting, or fluorescence in situ hybridization (FISH) Figure. An example of FISH-treated metaphase chromosomes Here, chromosomes 1, 2, and 4 were labeled yellow with FISH and the other chromosomes were stained red. Translocations between yellow and red chromosomes are detected. The left picture represents a normal cell (the numbers in the figure indicate chromosome numbers) and the right picture is an example of reciprocal translocation with two bi-color chromosomes (indicated by two arrows). A translocation break (t4:?) 2 2 4 1 1 t4 t4 Detecting Aberrations . Figure 1. Genomic region of the X chromosome containing the X;2 translocation breakpoint and TM4SF2 (Xp11.4 tetraspanin). a , Physical map of the Xp11.4 locus and genomic structure of TM4SF2 . The position of the breakpoint is indicated by a vertical dotted line. The eight exons are indicated (filled boxes) and their mapping showed that TM4SF2 spans at least 130 kb. STSs and ESTs are indicated (top) and the order of ESTs that map in the overlapping region between YAC clones 770D12 and 957F5 was defined here through their position on the BAC contig.
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