BIOL484 Study Guide - Midterm Guide: Agarose Gel Electrophoresis, Restriction Digest, Chromosome
6 Apr 2018
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Document Summary
The first step in mapping a mutant gene is to assign the gene to a broad region of one chromosome. The method presented here uses 48 snps evenly spaced along the chromosomes, allowing the whole genome to be scanned simulta- neously. The 48 primer sets are designed to amplify under identical conditions, and the snps are detected using identical restriction digestion conditions. The primers are arrayed in a 96-well plate to maximize reproducibility and to mini- mize set-up time. Two dna templates are generated, one from a mixture of 50 homozygous mutant animals and a control template from nonmutant animals (or an equal mixture of n2 and cb4856 animals). Unlinked snps should have equal n2 and cb4856 content. Thus, mutant templates and control templates should produce an equal proportion of n2 and cb products at unlinked loci. Linkage to a snp will result in an increase in the ratio of n2 dna/cb dna in the mutant relative to the control.
