2017整理分子生物学研究进展05 Biochip1029.ppt

Presentation outline Presentation outline Presentation outline Microarrays enable simultaneous measurement of the expression levels of thousands of genes in a sample Microarray: Glass slide with a matrix of thousands of spots printed on to it Each spot contains probes which bind to a specific gene 用于基因转录水平的检测、基因组分析和后基因组研究 美国斯坦福大学Davis等用PCR法从外周血淋巴细胞cDNA文库中得到了1046种cDNA片段，制成芯片。然后与热处理的T淋巴细胞和未处理的T细胞种提取的mRNA反转录出的单链cDNA片段杂交。经激光共聚焦扫描系统分析，共发现17个差异表达的基因，其中11个是被热诱导的，6个是被热抑制的。经序列分析证实，其中3个是未报导的新基因。 Presentation outline Complementary uses of DNA and tissue arrays DNA microarrays for gene expression Used to find biomarkers Relatively few tumor samples but thousands of markers (genes) TMAs for protein expression patterns Used to validate biomarkers large number (hundreds) of tumors, relatively few markers TMAs often used for validating tumor genes in cancer research The current classification systems of tumors are partially predictive of outcomes, and are based primarily upon morphologic criteria (grade, stage) The hope is that tumor markers may lead to improved diagnostic, prognostic and therapeutic applications in the clinic.   Tissue Array Section Real picture: brown staining Ki-67 Expression in Kidney Cancer Multiple measurements per patient:Several spots per tumor sample and several “scores” per spot Properties of TMA Data Highly skewed, non-normal, semi-continuous. Often a good idea to model as ordinal variables with many levels. Staining scores of the same markers are highly correlated Presentation outline Basic knowledge of flow cytometry BD CBATM液态芯片（科研）试剂盒（美国 2001） Luminex 100?液态芯片（科研）试剂盒（美国2004） FlowChipTM流式芯片试剂盒（西安微通2007） ? 临床诊断 ? 免疫监测 ? 药物研发 DNA Microarray Analysis Basic idea of Microarrays Gene expression Gene disease relation Gene-gene interaction Finding Co-Regulated Genes Understanding Gene Regulatory Networks … DNA Microarray Analysis One gene at a time before, now tens of thousands simultaneously DNA Microarray Analysis Microarrays desig