MCB 2050 Lecture Notes - Lecture 4: Tandem Repeat, Southern Blot, Contig
4 Sep 2018
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Mcb*2050 molecular biology of the cell topics 2. 1-2. 2: genomics. Less condensed chromosomes are, the more bands appear. Regions (at- rich) dna and relatively gene- poor, stain more darkly in g- banding (more distinct in prophase than metaphase) Specific gene visible via in situ hybridization, using either radioactive dna probes (detectable by photographic film) Exhibits visible chromosomal characters s such as reciprocal translocation of the philadelphia chromosome (9:22) Dna-biotin probe to avidin conjugated to a yellow fluorescent dye. Used to quantitatively distinguish different regions of the chromosome (telomere, centromere, arm specific probes) Microscopy of chromosome, staining bands, e. g. giemsa (g) or quinacrine (q) differential staining (banding pattern of stained chromosomes) Hybridize biotin- bound dna probe to cells on a glass slide. Incubate with avidin bound to a fluorescent dye. Mitotic chromosome smear, treat with the protease trypsin, stain with giemsa, observe by light microscopy. Hybridize radioactive (e. g. 32p) or fluorescent dna probe to metaphase cells on a glass slide.
