Biology 1002B Lecture Notes - Lecture 10: Sanger Sequencing, Shotgun Sequencing, Dna Microarray
29 Aug 2016
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Class 20: current trend in cost of dna sequencing. Very technological, cheap, easy to complete: steps in typical dna sequencing methodologies. Purification, fragmentation, amplification, sequencing fragments, assembly of fragmented pieces: structural features of dideoxynucleotides. They do not have a 3"- oh end, so it terminates dna elongation. Also has a fluorescent dye (can tell which base is where: basic mechanism of classical vs. automated sanger sequencing. Sequence fragments by incorporating dideoxynucleotides into 4 beakers (one base in each), run on gell, fluoresce with colour and identify which bases are where (shorter, travels further, ealier in the sequencing fragment) Different bases have different colour, all mix together with fragmented pieces, laser shoots through cylindrical gel bottom to top, identify which bases where (bottom, earlier: basic mechanism of pyrosequencing. Adapter binds to end of ds dna, ds dna split into ss dna, adapters bind to beads.
