Illumina-測序的原理和應(yīng)用課件

Illumina測序的原理和應(yīng)用

1精選ppt2021最新Illumina測序流程DataAnalysis

數(shù)據(jù)分析Sequencing

測序ClusterGeneration

簇生成LibraryPreparation

文庫制備測序流程2精選ppt2021最新AdapterandLibrary接頭和文庫接頭：與flowcell結(jié)合區(qū)Read1和Read2測序引物結(jié)合區(qū)Index

用來區(qū)分不同樣本3精選ppt2021最新Illumina測序流程DataAnalysis

數(shù)據(jù)分析Sequencing

測序ClusterGeneration

簇生成LibraryPreparation

文庫制備測序流程4精選ppt2021最新什么是flowcell？Flowcell即流動(dòng)槽，如載玻片大小，有8個(gè)通道即8條lane。5精選ppt2021最新1.SingleDNAlibrariesarehybridizedtoprimerlawnandextendedbypolymerase6精選ppt2021最新2.Double-strandedmoleculeisdenaturedandoriginaltemplateiswashedaway7精選ppt2021最新3.Newsynthesizedstrandiscovalentlyattachedtoflowcellsurfacetoformabridgeandextendedbypolymerase4.Double-strandbridgeisformed8精選ppt2021最新5.Double-strandbridgeisdenaturedandformtwocopiesofcovalentlyboundsingle-strandedtemplates6.Bridgeamplificationcyclerepeat9精選ppt2021最新MultiplebridgesareformeddsDNAbridgesaredenaturedandreversestrandsarecleaved10精選ppt2021最新9.Cleaved

reversestrandsarewashedawayandleavingaclusterwithforwardstrandsonly在Illumina的測序過程中，無論是單端測序還是雙端測序，都會(huì)用到特異選擇性鏈切斷的過程。其中單端測序的要切斷1次，雙端測序中的兩條鏈要先后各切斷1次（共2次）。單端測序：高碘酸希夫反應(yīng)。雙端測序：Illumina巧妙地利用了“甲酰胺基嘧啶糖苷酶（Fpg）”對(duì)“8-氧鳥嘌吟糖苷（8-oxo-G）”的選擇性切斷作用。11精選ppt2021最新Illumina測序流程DataAnalysis

數(shù)據(jù)分析Sequencing

測序ClusterGeneration

簇生成LibraryPreparation

文庫制備測序流程12精選ppt2021最新Hiseq2500Twoflowcell13精選ppt2021最新10.

Sequencingprimerishybridizedtoadaptersequence測序引物3‘端5‘端14精選ppt2021最新11.Onlyonenucleotideisaddedeverycycleasthe3’endisblocked15精選ppt2021最新12.Cleavethefluorandfreethe3’endthenrepeatabovestepsfornextcycle16精選ppt2021最新13.Sequencedstrandisstrippedoffandtheindexprimerisaddedfortoseqindex3‘端5‘端Indexprimer17精選ppt2021最新14.Repeatbridgeamplificationtoformreversestrandafterindexsequencing15.Sequencingthereversestrands

18精選ppt2021最新高通量測序的應(yīng)用DNA水平RNA水平基因組denovo、重測序外顯子捕獲測序DNA甲基化測序Chip-Seq16s、宏基因組轉(zhuǎn)錄組測序數(shù)字表達(dá)譜DGESmallRNAlncRNA19精選ppt2021最新高通量測序的應(yīng)用RNADNAProteinsRNALong

ncRNA轉(zhuǎn)錄組DGE原核真核有參無參比較轉(zhuǎn)錄組BSA-seq甲基化Ch