BIOL 2030 Lecture Notes - Lecture 27: Frederick Sanger, Thermal Cycler, Polymerase Chain Reaction
30 Mar 2017
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Lecture 27 - dna sequencing & polymerase chain. Two essential biochemical features for dna synthesis by dna polymerase. A short segment of double-stranded nucleic acid serves as a primer for initiation of dna synthesis. Dna is synthesized in the 5" to 3" direction o. Can only be elongated by the a primer. Both dna sequencing and the polymerase chain reaction requires dna synthesis in vitro. Single-stranded dna template: dna polymerase, dna primer, nucleotides. Chemical cleavage method by maxam and gilbert - published in 1977, but not commonly used now. Enzymatic dideoxy chain terminating method by fred sanger published in 1975 - very popular method and now semi-automated. Both strategies led to the nobel prize awarded in 1980. The oh between the 3" carbon and the oh on one of the phosphates (attached to another nucleotide) undergo a condensation reaction, allowing the two nucleotides to attach. Strategy for dna sequencing by the dideoxy chain termination method o.
