BIOC 2580 Study Guide - Final Guide: Motor Protein, Ribosomal Rna, Sanger Sequencing
16 Dec 2015
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Document Summary
Most data from one sequencing round in illumina. Dna template for synthesis produced by a plasmid placed in bacteria for replication. 4 separate reactions each with 4dntps and one of the ddntps fluorescently labelled, each with a different colour. Anytime a ddntp is incorporated, chain terminates resulting in dna fragments of different sizes and different labels. Strands separated by capillary electrophoresis by size so they each differ by one base and sequence is determined. Known sequence fragments added to the ends of the unknown fragment. Water droplets in oil contain a bead with an affixed. Each dntp washed over the plate one at a time. When a dntp is incorporated, protons are released. No bacterial cloning since all dna is amplified in vitro. Limit on number of bases that can be incorporated so shorter read lengths. Sequencing technologies page 1 detection, no electrophoresis a bead with an affixed primer, the single template annealed to the primer, dntps and a polymerase.
