BPS 3101 Study Guide - Midterm Guide: Gel Electrophoresis, Oligomer, Southern Blot
7 Mar 2015
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Oligomer-specific hybridization (oligomers for wild-type allele vs. mutant allele) In wt individuals (homozygous), only oligomers with 100% complementarity to wt sequence will give signals. In individuals with mutation (heterozygous), both oligomer types will give signals. For the homozygous mutant case, there only will be signal with oligomers that are 100% complementary to mutant sequence. Note for southerns, the sizes of hybridizing fragments will be depend on which oligomers & restriction enzyme you choose. For homozygous mutant, there would no signal with exon 2 probe. Note in northerns & rt-pcr experiments, the signals/product sizes will be correspondingly smaller (since exon 2 is not present). Model: inactivation of gene a if there is an is element in promoter region or within coding sequence except not if within c-terminal coding region. For longer pcr products, analyze restriction profiles (using s enzyme) to localize is element position. If is element near 3" end of gene, maybe no negative effect on protein function.
