BIOL 202 Lecture Notes - Lecture 15: Chromatography, Gene Duplication, Capillary Electrophoresis
Document Summary
Sanger: complementary addition of nucleotides by dna polymerase, ability to separate dna fragments by size, radiolabelling of nucleotides, terminal nucleotides (ddntp) No addition onto 3" site because of lack of hydroxyl group. * run 4 reactions in parallel (all the ddntps), run ge on each and the entire sequence can be predicted. Only one reaction required (improvement to original sanger sequencing) Capillary electrophoresis instead of a gel: fluorescence detector and a laser beam, producing a chromatogram (like in mimm 212) Next-gen sequencing: illumina/solexa like sanger, but with a reversible blocking nucleotide small primers bound to a piece of glass (coverslide) and instead of ddntps, a different kind of reversible nucleotide is added and gives of fluorescent signals. At each cycle of synthesis, nucleotide addition is blocked and then unblocked. Nucleotides added individually incorporation produces a flash of light. *contig: a sequence predicted from overlapping sequence reads n. b.
