1、第五部分：其他类型的芯片第五部分：其他类型的芯片 微缩芯片实验室（微缩芯片实验室（ Lab-on-a-chip) 药物运输芯片药物运输芯片 生理功能辅助芯片生理功能辅助芯片 纳米芯片纳米芯片Next Generation Sequencing Sample fragmentation Library preparation Sequencing reaction Data analysisRoche 454 焦磷酸测序焦磷酸测序Pyrophosphate SequencingIllumina Solexa 合成测序合成测序Sequence by SynthesizeABI SOLiD 连接法测
2、序连接法测序Sequence by Ligation第六部分第六部分 高通量测序技术简介高通量测序技术简介Roche 454 焦磷酸测序Pyrophosphate Sequencing基本原理454 sequencing： Emulsion PCR (emPCR)Mix DNA Libraryfluor removalODNAHNNOO3O5free 3 endXOH All 4 labelled nucleotides in 1 reactionSequencing-by-Synthesis (SBS)5GTCAGTCAGTCAGT35CAGTCATCACCTAGCGTAFirst base
3、 incorporatedCycle 1: Add sequencing reagentsRemove unincorporated basesDetect signalCycle 2-n: Add sequencing reagents and repeat1、每轮测序反应加入四种带有荧光标记的 dNTP，末端带有可以被去除的阻断基团2、每轮反应只能整合一个核苷酸，仪器读取相应的荧光信号3、信号读取结束，用化学方法去除阻断基团，进行下一轮测序反应1 2 3 7 8 94 5 6T T T T T T T G T T G C T A C G A T The identity of each base of a cluster is read off from sequential images根据每个点每轮反应读取的荧光信号序列，转换成相应的 DNA序列Base calling from the raw data
