Illumina测序的原理和应用 课件.pptx

Illumina 测序的原理和应用 ppt课件 Library Preparation 文库制备 Cluster Generation 簇生成测序流程 Sequencing 测序 Data Analysis 数据分析Illumina测序流程ppt课件接头：与flow cell结合区Read1和Read2测序引物结合区Index 用来区分不同样本Adapter and Library接头和文库ppt课件 Library Preparation 文库制备 Cluster Generation 簇生成测序流程 Sequencing 测序 Data Analysis 数据分析Illumina测序流程ppt课件什么是 flow cell？Flow cell 即流动槽，如载玻片大小，有8个通道即8条lane。ppt课件1. Single DNA libraries are hybridized to primer lawn and extended by polymeraseppt课件2. Double-stranded molecule is denatured and original template is washed awayppt课件3. New synthesized strand is covalently attached to flow cell surface to form a bridge and extended by polymerase4. Double-strand bridge is formedppt课件ppt课件5. Double-strand bridge is denatured and formtwo copies of covalently bound single-stranded templates6. Bridge amplification cycle repeatMultiple bridges are formeddsDNA bridges are denatured and reverse strands are cleavedppt课件9. Cleaved reverse strands are washed away and leaving a cluster with forward strands only在Illumina的测序过程中，无论是单端测序还是双端测序，都会用到特异选择性链切断的过程。其中单端测序的要切断1次，双端测序中的两条链要先后各切断1次（共2次）。单端测序：高碘酸希夫反应 。双端测序： Illumina巧妙地利用了“甲酰胺基嘧啶糖苷酶（Fpg）”对“8-氧鸟嘌吟糖苷（8-oxo-G）”的选择性切断作用。ppt课件 Library Preparation 文库制备 Cluster Generation 簇生成测序流程 Sequencing 测序 Data Analysis 数据分析Illumina测序流程ppt课件Hiseq2500Two flow cellppt课件3‘端测序引物5‘端10. Sequencing primer is hybridized to adapter sequenceppt课件11. Only one nucleotide is added every cycle as the 3’ end is blockedppt课件12. Cleave the fluor and free the 3’ end then repeat above steps for next cycleppt课件13. Sequenced strand is stripped off and the index primer is added for to seq index3‘端Index primerppt课件5‘端14. Repeat bridge amplification to form reverse strand after index sequencing15. Sequencing the reverse strands ppt课件基因组denovo、重测序外显子捕获测序DNA甲基化测序Chip-Seq16s、宏基因组DNA水平转录组测序数字表达谱DGESmall RNAlncRNARNA水平高通量测序的应用ppt课件高通量测序的应用RNAChip-seqDNA甲基化真核无参BSA-seq原核有参比较转录组ProteinLong ncRNA转录组DGEsRNAppt课件 Thank you!ppt课件高碘酸希夫反应：在合成DNA引物链的时侯，加入一个二醇基团，也就是相邻的两个碳各接一个羟基。在做完桥式