0.生物标志物(Biomarker)) 使用到抗体的高通量组学技术？ 核酸甲基化检测 结构基因组 基因组水平 -NGS 转录调控基因组. -MALDI-TOF 抗体芯片 -LC/MS OlinK(PEA)等标记技术 Simoa等数字ELISA技术 蛋白质组水平 -array 相互作用组学 蛋白修饰组学
使用到抗体的高通量组学技术？ 基因组水平 核酸甲基化检测 结构基因组 转录调控基因组. 抗体芯片 OlinK(PEA)等标记技术 Simoa等数字ELISA技术 相互作用组学 蛋白修饰组学. 蛋白质组水平 -MALDI-TOF -LC/MS 0.生物标志物（Biomarker） -NGS -array
I.DNA Microarray DNA Microarray? Scientists used to be able to perform genetic analyses of a few genes at once.DNA microarray allows us to analyze thousands of genes in one experiment! Prepare cDNA Probe Prepare Microarray "Normal" Tumor 交会入 绕 RT/PCR Label with Amounts microarray SCAN Microarray Technology
DNA Microarray? • Scientists used to be able to perform genetic analyses of a few genes at once. DNA microarray allows us to analyze thousands of genes in one experiment! I. DNA Microarray
I.DNA Microarray Purposes So why do we use DNA microarray? To measure changes in gene expressionlevels-two samples'gene expression can be compared from different samples,such as from cells of different stages of mitosis. To observe genomic gains and losses.Microarray Comparative Genomic Hybridization(CGH) To observe mutations in DNA
Purposes • So why do we use DNA microarray? – To measure changes in gene expression levels – two samples’ gene expression can be compared from different samples, such as from cells of different stages of mitosis. – To observe genomic gains and losses. Microarray Comparative Genomic Hybridization (CGH) – To observe mutations in DNA. I. DNA Microarray
I.DNA Microarray Usually made commercially. The Plate Made of glass,silicon,or nylon. Each plate contains thousands of spots,and each spot contains a probe for a different gene. A probe can be a cDNA fragment or a synthetic oligonucleotide,such as BAC (bacterial artificial chromosome set). Probes can either be attached by robotic means, where a needle applies the cDNA to the plate, or by a method similar to making silicon chips ■感E for computers.The latter is called a Gene Chip. GeneChip affymetrix Gonome Vede thma的SN把Nay年0 Biology for a better world 8P 利材2过设 Lg期40D E Dwe 1o1e
The Plate • Usually made commercially. • Made of glass, silicon, or nylon. • Each plate contains thousands of spots, and each spot contains a probe for a different gene. • A probe can be a cDNA fragment or a synthetic oligonucleotide, such as BAC (bacterial artificial chromosome set). • Probes can either be attached by robotic means, • where a needle applies the cDNA to the plate, • or by a method similar to making silicon chips • for computers. The latter is called a Gene Chip. I. DNA Microarray
I.DNA Microarray Let's perform a microarray! 1) Collect Samples. 基因组DNA 基因组DNA 秒照样本 实脸样本 2) Isolate mRNA. 3) Create Labelled DNA. 4)Hybridization. 参照样本标记 实验样本标记 Cy3(级色荧光) 5) Cy5(红色黄光 Microarray Scanner. 6) Analyze Data. 两个样品共同 杂交一张芯片 laser ratio Sample Purification RT Coupling Hybridization Scanning Normalization and washes and analysis
Let’s perform a microarray! 1) Collect Samples. 2) Isolate mRNA. 3) Create Labelled DNA. 4) Hybridization. 5) Microarray Scanner. 6) Analyze Data. I. DNA Microarray