PHYSICS 102 Lecture Notes - Lecture 27: De Novo Synthesis, Polyacrylamide Gel Electrophoresis, Fluorescent Tag
Document Summary
Identification of disease-associated mutations: sequencing to find this. Identification of genetic polymorphisms: genome sequencing of micro-organisms for identification of novel drug targets etc, express novel proteins, design drugs and test them etc. Controlled de novo synthesis of dna fragments from a defined origin: enzyme based. Termination at specific random points along target sequence. Templates can be ss dna, ds dna plasmids, pcr products etc. Fluorescent analogues and automation of dna sequencing: reaction can be carried out in 1 tube as termination is identified by colour. Dye terminator dideoxy sequencing: detect different fluorescent tag products as they pass detector, which fluorophore is with what base. If a deoxynucleotide is incorporated the polymerisation reactions continues as there is a 3" hydroxyl group on the end so a base can be added to form newly synthesised piece of dna. If a dideoxynucleotide is incorporated the polymerisation reaction terminates as dna pol doesn"t have a 3" hydroxyl to add any bases.
