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Unformatted text preview: 1 Gene Expression & DNA Microarrays Background and basis for approach Array Technologies First published example (from yeast) Bioinformatics and Array Data Analysis Applications of Expression Profiling Human cells Other uses of microarrays 1 Concept: use DNA microarrays to study expression of all genes in the genome 1. Genome sequences became available (1990s-present) Yeast ( S. cerevisiae ) first then fly, worm, human 2. High throughput methods for analysis demonstrated: robotics, bioinformatics 2 Basis of method: Genes are expressed at different levels RNA gives us a way to measure gene regulation 3 mRNA can be converted into cDNA and tagged (Labeled) with Fluorescent Molecules Reverse Transcriptase mRNAs Fluorescent-labeled cDNAs 4 DNA Bound to a Solid Substrate Can Be Used to Detect Tagged DNA Fluorescent-labeled cDNAs DNA array 5 The Number of Fluorescent Tags at Each Spot Indicates Abundance Level Fluorescent-labeled cDNAs 2 6 Spotted DNA Arrays Mechanical deposition of liquid onto coated glass slides (DNA of each gene) Capable of spotting any nucleic acid (or protein) Max. features: ~ 40,000 per slide Each spot represents an individual gene 7 28,000 DNA Sequences 8 9 10 Overview of method Step 1: Produce microarrays: PCR amplify yeast ORFs Use robot to spot PCR products on glass slide Step 2: Perform experiment with yeast cells...
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This note was uploaded on 06/11/2009 for the course MIC 170 taught by Professor Shiozaki during the Spring '09 term at UC Davis.
- Spring '09