DNA Microarrays for Statisticians. cDNA Microarray Applications Revolutionized gene expression studies Diagnose cancer Expression profile of tumor progression.

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Presentation transcript:

DNA Microarrays for Statisticians

cDNA Microarray Applications Revolutionized gene expression studies Diagnose cancer Expression profile of tumor progression Discriminate types of cancers Track time course of gene expression in sporulating yeast Track cell-cycle genes in synchronized yeast Hopefully determine more targets for E2F

Making a cDNA Microarray Critical parameters: Spot size Spot shape DNA concentration 384 Well Plates …..24 A…P Wash Technologies: Pins or Needles Pin and Ring Inkjet Printer

Target Generation Sense Anti Sense 5’ TTTTT AAAAA 3’ Reverse Transcriptase DNA Polymerase RNA DNA

A Yeast Microarray

Affymetrix Photolithography

Experimental Protocol Isolate total RNA from 8-10hr old embryos Make cDNA Make biotinylated cRNA Hybridize 16 hours at 45C Wash and stain Scan Analyze(generate average difference for each probe set)

Target Generation Sense Anti Sense 5’ TTTTT AAAAA T7 comp primer 3’ Reverse Transcriptase DNA Polymerase RNA polymerase RNA DNA RNA