BIOL240 Lecture Notes - Lecture 10: Proteome, Reverse Transcriptase, Polyacrylamide Gel Electrophoresis

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The genome is the entire complement of genetic information which includes genes, regulatory sequences, and non-coding sequences. Genomics is the discipline involving mapping, sequencing, analyzing, and comparing genomes. Functional genomics is assigning function to unknown genes by analyzing biochemical and physiological effects of mutants. Sanger/dideoxy dna sequencing is the classic method of sequencing. There are three main steps to this process. This entire process is based on the chemistry of dideoxynucleotides. Dna polymerase requires a free 3" oh to continue synthesis. So you basically do this in 4 separate tubes with dideoxygtp, ddatp, ddttp, and ddctp. Once you do gel electrophoresis, you can read the exact order (the smaller the fragment the faster it will run) but you have to read it backwards. All the c"s are radioactively marked with p52. We"ve updated this version to make all 4 dideoxyribonucleotides in one tube but all with a different colour fluorescence marker, again with gel electrophoresis or a laser signal.

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