BIOL201 Lecture Notes - Dapi, Interphase, Centromere
11 Apr 2014
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Fluorescence in situ hybridization: how to fish, overview. Identify genes or chromosomes amongst interphase or metaphase chromosomes. Make a probe with a random prime labeling reaction one or more bac clones fluorescently labeled nucleotide. Put some cells onto a slide, denature the dna, add the probe. Example 1: cell = amniotic (fetal) cell blue (dapi) = dna green = chr 13 cen red = chr 21 cen. Example 2 cell = amniotic blue = dna light blue = chr 18 cen green = x chr cen red = y chr cen. Maternal age effect for trisomy-21: fish with locus specific probes. Identify deletions and duplications cells = child blue = dna green = 15 cen red = 15q11 and 15q23. 9 cells = child blue = dna white = chr 1 purple = chr 9. How many different chromosomes do humans have: 20, 21, 22, 23, 24. Step 3 use six filters to take six images.
