Affymetrix基因芯片产品培训课件tum.pptx

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1、AFFYMETRIX基因芯片产品 Up to Up to 7,007,000,000 0,000 featuresfeatures /chipchip5-11m5-11mRNA targetRNA targetThousands of copies of a specificThousands of copies of a specificoligonucleotide probe in oligonucleotide probe in each featureeach feature Raw data image with 1-Raw data image with 1-6.96.9 mil

2、lion differentmillion differentcomplementary probescomplementary probes OligonucleotideOligonucleotideprobeprobe*1.28 cm1.28 cmHybridized probe cellHybridized probe cell*5”5”5”5”芯片产品分类表达谱芯片-mRNA3IVTWTMicroRNA芯片lncRNA芯片SNP/CNV芯片甲基化&转录因子芯片表达谱芯片3IVTHuman U133 Plus 2.0Mouse U430 2.0Rat U230 2.0PrimeView

3、&Almac XcelWTHuman/Rat/Mouse Gene1.0 STHuman/Rat/Mouse Exon1.0 STHTA2.0表达谱芯片探针设计Exon 1.0 ST基因组DNA初始转录本RNA可变剪切转录本Gene 1.0 ST3端表达谱表达谱芯片实验步骤样本RNA抽提&质检cDNA合成及片段化生物素标记芯片杂交-洗脱-扫描芯片数据标准化GENE系列芯片TranscriptcoverageofthearrayHumanGene1.0STArrayHumanGene2.0STArrayTotalRefSeqtranscriptscovered36,07940,716NM?Ref

4、Seqcodingtranscript,well-establishedannotation32,02030,654NR?RefSeqnon-codingtranscript,wellestablishedannotation2,9675,638XM?RefSeqcodingtranscript,provisionalannotation579996XR?RefSeqnon-codingtranscript,provisionalannotation5133,428lincRNAtranscripts46611,086RefSeq(Entrez)genecount21,01424,838EXO

5、N系列芯片DesignStatisticsSummaryProbeSets1.4millionExonClusters1millionSupportedbyPutativeFull-LengthmRNA289,961probesetsSupportedbyEnsemblTranscripts306,583probesetsSupportedbyEST665,175probesetsSupportedbyMouseorRatmRNA220,262probesetsSupportedbyGenePrediction883,105probesetsProbeSelectionRegionAlongt

6、heentirelengthofthetranscriptsProbes/ProbeSelectionRegion4BackgroundSubtractionStrategyMedianintensityofupto1,000backgroundprobeswiththesameGCcontentTotalFeaturesPerArray5,500,000InterrogatedStrandSenseHTA2ArraycontentProteincodingcontentNon-proteincodingcontentGenes(transcriptclusters)44,69922,829T

7、ranscripts245,34940,914Exons560,472109,930Exonclusters296,05882,444Splicejunctions(probesets)339,1460表达谱芯片应用U133经典，相对旧，无价格优势，不做推荐Gene系列基本，仅满足一般表达量检测需求Exon系列全面，精确表达量检测HTA2最佳，最全&最精确表达量EG1.0最佳，唯一能够获得转录本表达量MICRORNA芯片MicroRNA 4.0microRNAPre-microRNAsnoRNA&scaRNAMICRORNA芯片实验步骤样本RNA抽提&质检Poly(A)加尾&FlashTagB

8、iotinHSR连接生物素标记芯片杂交-洗脱-扫描芯片数据标准化MICRORNA4.0特点100%覆盖miRBase V20包括所有数据库记录的物种人 2578小鼠 1908大鼠 728重复性0.95技术重复生物重复LNCRNA芯片Human/Mouse/Rat Gene2.0HTA2.0(Human)EG1.0-Human/MouseLNCRNA芯片实验流程样本RNA抽提&质检cDNA合成及片段化生物素标记芯片杂交-洗脱-扫描芯片数据标准化SNP/CNV芯片SNP6.0CytoScan 750KCytoScan HDOncoScan订制芯片CYTO芯片适用样本细胞组织血液唾液骨CYTO HD

9、270万CNV探针75万SNP探针特异性灵敏度高覆盖率1 marker/1-3KB1marker/5KB高性能高分辨率36000120005005004000035000300002500020000150001000050000RefseqOMIMSangerISCA基因数量CYTO系列芯片价值杂合性缺失单亲二倍体三倍体样本异质性家系分析甲基化芯片 Human Promoter1.0 RDNA甲基化CpG岛在CpG甲基化结合蛋白和DNA甲基化转移酶的作用下，使CpG二核苷酸5端的胞嘧啶转变成为5甲基胞嘧啶。通常是基因的启动子区域，5端非翻译区和第一个外显子区，CpG序列密度非常高，超过均值5倍以上，为鸟嘌呤和胞嘧啶的富集区。研究甲基化意义DNA甲基化过低CpG岛高甲基化出生后发育肿瘤整体甲基化水平发育衰老样品交联染色质片段化CHIP洗涤纯化PCR扩增DNA片段化标记杂交洗脱扫描PROMOTER1.0R实验流程谢谢观看/欢迎下载BY FAITH I MEAN A VISION OF GOOD ONE CHERISHES AND THE ENTHUSIASM THAT PUSHES ONE TO SEEK ITS FULFILLMENT REGARDLESS OF OBSTACLES.BY FAITH I BY FAITH