Illumina测序的原理和应用-课件

Illumina测序的原理和应用

1ppt课件Illumina测序的原理和应用Illumina测序流程DataAnalysis

数据分析Sequencing

测序ClusterGeneration

簇生成LibraryPreparation

文库制备测序流程2ppt课件Illumina测序流程DataAnalAdapterandLibrary接头和文库接头：与flowcell结合区Read1和Read2测序引物结合区Index

用来区分不同样本3ppt课件AdapterandLibrary接头和文库接头：Illumina测序流程DataAnalysis

数据分析Sequencing

测序ClusterGeneration

簇生成LibraryPreparation

文库制备测序流程4ppt课件Illumina测序流程DataAnal什么是flowcell？Flowcell即流动槽，如载玻片大小，有8个通道即8条lane。5ppt课件什么是flowcell？Flowcell即流动槽，1.SingleDNAlibrariesarehybridizedtoprimerlawnandextendedbypolymerase6ppt课件1.SingleDNAlibrariesare2.Double-strandedmoleculeisdenaturedandoriginaltemplateiswashedaway7ppt课件2.Double-strandedmolecule3.Newsynthesizedstrandiscovalentlyattachedtoflowcellsurfacetoformabridgeandextendedbypolymerase4.Double-strandbridgeisformed8ppt课件3.Newsynthesizedstrandis5.Double-strandbridgeisdenaturedandformtwocopiesofcovalentlyboundsingle-strandedtemplates6.Bridgeamplificationcyclerepeat9ppt课件5.Double-strandbridgeisdeMultiplebridgesareformeddsDNAbridgesaredenaturedandreversestrandsarecleaved10ppt课件Multiplebridgesareformed10p9.Cleaved

reversestrandsarewashedawayandleavingaclusterwithforwardstrandsonly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。11ppt课件9.CleavedreversestrandsarIllumina测序流程DataAnalysis

数据分析Sequencing

测序ClusterGeneration

簇生成LibraryPreparation

文库制备测序流程12ppt课件Illumina测序流程DataAnalHiseq2500Twoflowcell13ppt课件Hiseq2500Twoflowcell13ppt课件10.

Sequencingprimerishybridizedtoadaptersequence测序引物3‘端5‘端14ppt课件10.Sequencingprimerishybr11.Onlyonenucleotideisaddedeverycycleasthe3’endisblocked15ppt课件11.Onlyonenucleotideisad12.Cleavethefluorandfreethe3’endthenrepeatabovestepsfornextcycle16ppt课件12.Cleavethefluorandfree13.Sequencedstrandisstrippedoffandtheindexprimerisaddedfortoseqindex3‘端5‘端Indexprimer17ppt课件13.Sequencedstrandisstrip14.Repeatbridgeamplificationtoformreversestrandafterindexsequencing15.Sequencingthereversestrands

18ppt课件14.Repeatbridgeamplificat高通量测序的应用DNA水平RNA水平基因组denovo、重测序外显子捕获测序DNA甲基化测序Chip-Seq16s、宏基因组转录组测序数字表达谱DGESmallRNAlncRNA19ppt课件高通量测序的应用DNA水平RNA水平基因组denovo、重测高通量测序的应用RNADNAProteinsRNALong

ncRNA转录组DGE原核真核有参无参比较转录组BSA-seq甲基化Chip-seq20ppt课件高通量测序的应用RNADNAProteinsRNALong

Thankyou!21ppt课件21ppt课件Illumina测序的原理和应用

22ppt课件Illumina测序的原理和应用Illumina测序流程DataAnalysis

数据分析Sequencing

测序ClusterGeneration

簇生成LibraryPreparation

文库制备测序流程23ppt课件Illumina测序流程DataAnalAdapterandLibrary接头和文库接头：与flowcell结合区Read1和Read2测序引物结合区Index

用来区分不同样本24ppt课件AdapterandLibrary接头和文库接头：Illumina测序流程DataAnalysis

数据分析Sequencing

测序ClusterGeneration

簇生成LibraryPreparation

文库制备测序流程25ppt课件Illumina测序流程DataAnal什么是flowcell？Flowcell即流动槽，如载玻片大小，有8个通道即8条lane。26ppt课件什么是flowcell？Flowcell即流动槽，1.SingleDNAlibrariesarehybridizedtoprimerlawnandextendedbypolymerase27ppt课件1.SingleDNAlibrariesare2.Double-strandedmoleculeisdenaturedandoriginaltemplateiswashedaway28ppt课件2.Double-strandedmolecule3.Newsynthesizedstrandiscovalentlyattachedtoflowcellsurfacetoformabridgeandextendedbypolymerase4.Double-strandbridgeisformed29ppt课件3.Newsynthesizedstrandis5.Double-strandbridgeisdenaturedandformtwocopiesofcovalentlyboundsingle-strandedtemplates6.Bridgeamplificationcyclerepeat30ppt课件5.Double-strandbridgeisdeMultiplebridgesareformeddsDNAbridgesaredenaturedandreversestrandsarecleaved31ppt课件Multiplebridgesareformed10p9.Cleaved

reversestrandsarewashedawayandleavingaclusterwithforwardstrandsonly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。32ppt课件9.CleavedreversestrandsarIllumina测序流程DataAnalysis

数据分析Sequencing

测序ClusterGeneration

簇生成LibraryPreparation

文库制备测序流程33ppt课件Illumina测序流程DataAnalHiseq2500Twoflowcell34ppt课件Hiseq2500Twoflowcell13ppt课件10.

Sequencingprimerishybridizedtoadaptersequence测序引物3‘端5‘端35ppt课件10.Sequencingprimerishybr11.Onlyonenucleotideisaddedeverycycleasthe3’endisblocked36ppt课件11.Onlyonenucleotideisad12.Cleavethefluorandfreethe3’endthenrepeatabovestepsfornextcycle37ppt课件12.Cleavethefluorandfree13.Sequencedstrandisstrippedoffandtheindexprimerisaddedfortoseqindex3‘端5‘端Indexprimer38ppt课件13.Sequencedstrandisstrip14.Repeatbridgeamplificationtoformreversestrandafterindexsequencing15.Sequencingthereversestrands

39ppt课件14.Repeatbridgeamplificat高通量测序的应用DNA水平RNA