Chemical and Biochemical Engineering 2290A/B Lecture Notes - Lecture 12: Hybridization Probe, Shotgun Sequencing, Metagenomics
20 Apr 2016
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Document Summary
Genomics; study of molecular organization ; structural(physical nature), functional(interactions), and comparative. Sanger method**; chain-termination dna sequencing method by using ddntp"s (reaction stopping nucleotides) Need: single strands of dna with primer, dna polumerase i, 4 deoxynucleotides, and small amount of each ddntp(labled) Random insertion of ddnto generates dna fragments of different lengths. Gell imaged and sequence read bottom to top, small fragments migrate the furthest. Automated sequencing(shotgun sequencing): labeled with colored fluorescent dye, read by a laser. Analysis of genome data using computers-structure, content and arrangement. Uses annotation to determine the location of genes. Identifies open reading frame (orf) in the genome. Provides info on metabolic pathways, transport mechanisms, and regulatory and signal transduction mechanisms. Gene chips; determine which genes are expressed at a specific time. Each dna spot(probe) represents a single gene of orf. Robotic application of dna probe, expressed sequence tags for eucaryotes: photolithography. Analysis of the gene expression: targets are labled with fluorescent.
