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Lecture 13

biol202 - lecture 13

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PHGY 210
John White

ASO -short enough to allow differential hybridization -long enough to insure locus specifity At any place in the genome, what are the odds that nucleotide A is in that specific place? Sickle-cell anemia oligonucleotide that encompasses mutation normal allele only hybridizes only when Hg A is there mutated allele only hybridizes when there’s Hg S Blue allele has mutated form Taken DNA samples from individuals, put on 2 separate pieces of cellulose (both membrances0 !st individual is heterozygous, put on both membranes Hybridization with ASO, remove excess probe Homo: hyb = black dot -normal allele hyb. w/ normal probe -mutated allele hyb/ w/ mutated probe Hetero: can hyb. w/ both probes Cystic Fibrosis with ASO Mutation that removes three nucleotides is in open reading frame (remove one amino acid) > mutated form of sequence -mutated form will have same sequence except for three sequences -mutated can hyb. w/ normal ASO and mutated ASO Screen blood samples from people for the presence of sickle-cell allele) presence of diagnostic T rather than the normal A) -Want to get single cell allele A. view RBCs under microscope and look for sickle shapes -cannot see allele under microscope B. Denature DNA in blood samples and perform ASO hybridization screen C. Determine DNA sequence of each allele using… as oligo primer, thereby obtaining sequence information over Primer contains the mutation -won’t get mutation, first nucleotide will always have mutation (don’t know if it doesn’t work of if it’s a normal allele) -has to be outside -won’t work for heterozygotes LECTURE 12: Large scale chromosomal changes in plants Centromere Centromere separates short arm (p) from long arm (q) Short arm: “petit” Long arm: q is after p in the alphabet How to visualize Chromosomes • Grow WBS from blood sample • Stimulates cells to divide, increasing cells in mi
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