Data Mining of GeneChips® Devang Shah INT3470 Course Project March 14, 2002 Outline Background Molecular Biology 101 GeneChip® Technology Synthesis, Sample Preparation, Scanning Data Flow and Management Multidrug Resistance Preliminary Data Analysis Affymetrix Microarray Suite Scoring posite Analysis Statistical Algorithms (parison Analysis) Data Mining Tool (Preview) SOM Algorithm Correlation Coefficient Clustering Molecular Biology 101 A -> T G -> C Traditional methods result in “one gene at one time” analysis The need for higher throughput has led to the development of two (competing) technologies cDNA Spotted Array cDNA Microarrays utilize various technologies (spotting, piezoelectric, drop-touch) to create arrays of entire genomes using 500-5000 bp oligos representing entire genes Microarrays are scanned using laser microscopy with fluorescent labeled DNA GeneChip® Technology Utilizes photolithography binatorial chemistry to synthesize microarrays consisting of ~25mer oligos representing smaller regions of various genes Probe Cell (PM or MM) Probe Pair (PM + MM) Probe Set ( ~18 Pairs) Typical experiment involves isolation of mRNA, synthesis of cDNA, fragmentation, labeling , and hybridization Scan results in a raw data file (*.CHP) Data Management Nat 1999 Jan;21(1 Suppl):51-5 Gene expression informatics--it's all in your mine. Bassett DE Jr, Eisen MB, Boguski MS Use in Functional Genomics Antimicrobial resistance in bacteria Multi-drug pumps (MDR’s) Facilitate extrusion of amphipathic cations (toxic) TolC (E. coli), NorA (S. aureus) Evolutionary paradigm: Why do plants continue to produce pounds if they are extruded??? Perhaps they produce MDR inhibitors that bined with these normally ineffective antimicrobials provide a strong synergistic effect Methods E. coli TolC mutants were challenged with pounds for several hours and then assayed for gene expression S. aureus (WT + NorA Mutant) susceptibility to various plant pounds were test
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