MGY250H1 Study Guide - Final Guide: Cytokinesis, Okazaki Fragments, Allele

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Define hierarchical shotgun sequencing, dna cloning, sanger sequencing and understand their uses. Dna is randomly sheared and these fragments are then cloned into a bacterial host this is a. Contains overlapping pieces of dna, only some would be needed for a map. Take fragments further fragment into shotgun clone, sequence, and assemble the reads. To clone dna, the area of interest must be cut out with restriction enzymes. Vector/plasmid and the piece of dna are joined by ligation. Plasmid is transformed into bacteria results in recombinant dna. Recombinant dna is further purified from the host dna. Heat double stranded dna separation into single stranded dna. Test tube contain dntps and ddntps: ddntps prevent the dna from growing any further so you get fragments of different lengths, fragments are run on the gel, separated, and assembled. Uses: allowed the sequencing of large reads and understanding the genome. P arm = shorter arm, q arm = longer one.

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