第三讲利用基因芯片进行基因表达谱分析.pptVIP

Decoding Quality Metrics BeadArray Formats Sentrix Array Matrix Sentrix BeadChip Formats Human-6 HumanRef-8 Universal-16 Custom-16 Human-1 BeadStation?扫描系统 激光共聚焦扫描系统 高分辨率, 0.8 micron 双色激光 at 532 635 nm 超高分辨率 全自动化定位,扫描 BeadArray Reader Confocal laser scanning system Sub-micron, 0.8 micron Two lasers 532, 635 nm Supports Cy3 Cy5 imaging Auto-focusing, centering and tilting without photo bleaching Supports genotyping, gene expression applications Scans both Array Matrix and BeadChip microarrays Analytical Fluorescent Image * Five years ago microarray analysis was focused on gene expression profiling and some genotyping. Today scientists are exploring multiple applications that span both DNA and RNA analysis (mirroring the previous slide). aCGH is used to study chromosome alterations: deletions/duplications that occur in cancer and genetic disorders. ChIP-on-chip (Location Analysis) is used to study protein-DNA binding. When those proteins are transcription factors, LA bridges the genome, proteome and transciptome. There are at least three characteristics of a microarray platform that are important to extend into new applications. Greater density is important because, unlike gene expression profiling that queries 30,000 genes, scanning experiments in applications like aCGH and LA benefit from highest possible resolution. Splice variant analysis also benefits from higher numbers of feature. Sensitivity is important because applications like aCGH require consistent detection of small (2x) fold changes. And in many of these new apps, samples are limited, placing a premium on lowest limit of detection. Flexibility is important because in emerging applications (e.g. microRNA, splicing) new information must be incorporated into new microarray designs frequently. Flexibility also enables the kind of technology development required to perfect emerging applications (e.g. microRNA analysis). * * * * * Target Preparation RNA isolation is the first step. 1-2