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NextGenerationSequencing 454sequencing:EmulsionPCR(emPCR) ClonalSingleMoleculeArrays单分子克隆ReversibleTerminatorChemistry可逆终止反应Sequencing-by-Synthesis(SBS)1Solexa测序Workflow 文库制备:微珠单分子克隆1024种8碱基探针 4色荧光,4种双核苷酸,每色荧光有256个探针(4^6)连接法测序(一)连接法测序(二)连接法测序(三)Advantage&disadvantage高通量测序的应用Transcriptomeresequencing:TranscriptomecharacteristicsDigitalexpressionprofiling(1):人大脑组织与UHR(UniversalHumanReference)的表达差异Digitalexpressionprofiling& microRNAre-sequencing:ChIP-seq(1): 人一号染色体DNA-蛋白相互作用ChIP-seq(2):Methy-seq(1): 肿瘤和MCF7细胞系中BRCA!启动子区域的甲基化差异Somehighlights:CorrelationbetweenChIP-SeqandhispriorSAGE-likemethod(calledGMAT)hasr=0.906‘HowevertheresolutionwithChIP-Seqwasdramaticallyhigher.Furthermore,ChIP-Seqwasmoresensitiveandgeneratedlessfalse-negativeregions’12,726geneswhosetranscriptionlevelsareknowninCD4+T-cellswerecorrelatedwiththehistonemodificationsand35,961PolIIbindingsite‘islands’wereidentified‘Thiscost-effectivemethodproducesdigital-qualitydataandshouldfindbroadapplicationsinoureffortstounderstandthecontributionofthehumanepigenomesingeneexpressionandepigeneticinheritance’部分参考文献阅读藤晓坤,肖华胜*,基因芯片与高通量DNA测序技术前景分析。中国科学C辑:生命科学。2008:(38)10:1-9。 TengX,XiaoH.PerspectivesofDNAmicroarrayandnext-generationDNAsequencingtechnologies.SciChinaCLifeSci.2009;52(1):7-16