第二代高通量测序技术的简介.pptx

Next Generation Sequencing; ;454 sequencing： Emulsion PCR (emPCR);; ;Clonal Single Molecule Arrays单分子克隆;Reversible Terminator Chemistry可逆终止反应;Sequencing-by-Synthesis (SBS);1;Solexa 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;Transcriptome resequencing：;Transcriptome characteristics;Digital expression profiling（1）：人大脑组织与UHR（Universal Human Reference）的表达差异;Digital expression profiling microRNA re-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;Some highlights:Correlation between ChIP-Seq and his prior SAGE-like method (called GMAT) has r=0.906‘However the resolution with ChIP-Seq was dramatically higher. Furthermore, ChIP-Seq was more sensitive and generated less false-negative regions’12,726 genes whose transcription levels are known in CD4+ T-cells were correlated with the histone modifications and 35,961 Pol II binding site ‘islands’ were identified‘This cost-effective method produces digital-quality data and should find broad applications in our efforts to understand the contribution of the human epigenomes in gene expression and epigenetic inheritance’;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。 Teng X, Xiao H. Perspectives of DNA microarray and next-generation DNA sequencing technologies. Sci China C Life Sci. 2009 ;52(1):7-16 ;;Reversible Terminator Chemistry可逆终止反应;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);Advantage disadvantage;Advantage disadvantage;Some highlights:Correlation between ChIP-Seq and his prior SAGE-like method (called GMAT) has r=0.906‘However the resolution with ChIP-Seq was dramatically higher. Furthermore, ChIP-Seq was more sensitive and generated less false-negative regions’12,726 genes whose transcription levels are known in CD4+ T-cells were correlated with the histone modifications and 35,961 Pol II binding site ‘islands’ were identified‘This cos