MCB 2410 Lecture Notes - Lecture 18: Bacterial Artificial Chromosome, Whole Genome Sequencing, Abo Blood Group System
12 Apr 2018
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Document Summary
Nail-patella and abo blood type are linked: abo blood group locus maps to 9q34. Cftr found by positional cloning linkage analysis on families with cystic fibrosis: chromosome 7 markers linked with cr, more linkage narrows size of region of interest, clones from region isolated by chromosome walking and chromosome jumping. Northern blot analysis: evaluates rna expression in various tissues, rna gel membrane probed with cf candidate gene, hybridization means that rna is present in that particular sample, demonstrated linked cf gene expressed in affected tissues. Map based approach: use detailed genetic and physical maps to identify least number of clones to cover genome (minimum tiling path) then sequence and assemble that small number of clones, 2. Whole genome shotgun sequencing: shear entire genome into small pieces and clone, sequence millions of clones, e. g. 3. 2 billion bp/700 bp = 4. 5 million sequencing reactions, assemble all those sequences.
