SOCR 330 Lecture Notes - Pyrosequencing, Chain Termination, Functional Genomics
Document Summary
Most widely used sequencing methods today based on sanger sequencing. Synthesizing new dna strand using complementary strand of dna to be sequenced as template. Adding artificial pentose sugar (dideoxyribose) to reaction mixture to stop synthesis at random. Separating fragments of synthesized dna using gel electrophoresis. Uses pcr (polymerase chain reaction) to synthesize new strand of dna using unknown sequence at template. Instead of dideoxynucleotide incorporation and chain termination (as in sanger reaction), identifies last base added by pyrophosphate release. Pyrophosphate used to make atp which then provides energy for luciferase/luciferin reaction to generation chemoluminescence. Genomics = branch of genetics that studies whole genomes. Different types of dna sequence present in genome. Eukaryotic genomes often contain large quantities of non-expressed dna. Excess dna is present that does not seem to be essential to the development or evolutionary progression of eukaryotes. Due to presence of repeat nucleotide sequences within many genomes. Each sequence present only once in haploid genome.
