BIOL 202 Lecture Notes - Lecture 20: Deoxycytidine Triphosphate, Tata Box, Chromatography
5 Nov 2015
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February 20th, 2015: fred sanger (1974-1977, takes advantage of dna replication and tech at the time, complementary addition of nucleotides by dna polymerase, ability to separate dna fragments by size, radiolabelling of nucleotides, terminal nucleotides. Instead of doing 4 reactions, did 1 reaction. Dna still moves towards the positive pole through the capillary. There is a fluorescence detector and a laser. Get a reading of the amount of signal of each colour - chromatogram: a messy chromatogram with 2 peaks at one location could be due to a splice site mutation. Instead of a ddntp there is a reversible blocking nucleotide: 454/pyrosequencing. From sequences to genomes: the concept: cut up genomes into fragments, sequence each fragment, overlap sequence reads and try to align them to create contigs for a complete sequence, highly repetitive sequences are hard to align and sequence. coverage - sequencing errors occur, need many reads of each sequence to.
