04博士研究生课高级生物化学与分子生物学专题肿瘤功能基因组学许丽艳详解.pptVIP

DNA Recombination Technology Gene Chip Strategy of Functional Genomics RNA interference DNA Analysis Southern Blot: telomere length; amplification; deletion PCR: mutation; DNA methylation In situ hybridization: gene location; translocation DNA Chip: SNP Bioinformatics DNA Analysis Southern Blot: telomere length; amplification; deletion PCR: mutation; DNA methylation In situ hybridization: gene location; translocation DNA Chip: SNP Bioinformatics RNA Analysis Northern Blot : expressed genes RT-PCR: expressed genes In situ hybridization: gene location cDNA microarray : differentially expressed genes Bioinformatics Protein Analysis Western Blot: protein concentration Immunohistochemical Technique: protein location 2-D Electrophoresis: differerentially expressed proteins Bioinformatics Cover of Science 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 First Microarray Patent Issued 1st Catalog GeneChip? Product Roche EasyAccess Commercial launch West Sacramento 2002 Roche AmpliChipTM launched 2003 U133 Set 10K 1991 2004 100K Worlds First Diagnostic Microarray System Launched by Affy in Europe 11 μm Millions of copies of a specific oligonucleotide probe Image of Hybridized GeneChip Array 500,000 different complementary probes Single stranded, labeled ‘target’ Oligonucleotide ‘probe’ GeneChip? Array Hybridized Probe Cell 1.28cm 1.28cm Perfect Match PM MM Mismatch 独具创新的PM-MM探针设计 Oligo 探针 ATCGGTAGCCATGCATGAGTTACTA ATCGGTAGCCATCCATGAGTTACTA 13 MisMatch Base Discriminate against background signals Increasing sensitivity at low target concentrations DNA RNA Genotyping: Is it A or B? Gene Expression: Which genes? How much? Eukaryotic Cell One Genechip System DNA--Genotyping Family based linkage studies Whole-genome disease association Pharmacogenetics Chromosomal copy number Population genetics Linkage Disequilibrium Resequencing RNA--Expression Analysis Biomarker discovery Mechanism of Act