其他-rna-seq上練習(xí)從數(shù)據(jù)到功能分析

RNA-seqVSZhangetal.NatureReviewsGenetics,RNA-seq 1.Libraryconstructionwith 2.ClusterGenerationbybridge 3.Sequencingby 4.Massivelyparallel 5.Reads 6.Transcriptsabundance 7.Differentiallyexpressedgenes 8. NextGenerationSequencingLibraryMoreilluminaClusterGeneration:BridgePCR SequencingSequencingMultiplexingisachievedbyusingsamplespecificadaptorsinlibrarypreparationstep,thensequencingthemalltogetherinonelibraryPE=pairedend(mate

SE=singleIs2*50,000,000=MoreinformationonNewsplicevariantsNewstructuralvariationMoreperbasecoverageTypicallybetterma

LessinformationThe2ndfragmentisredundantinformationforcoveragepurposes.UsefulRSeQC:genebody 對各 fication單位的介紹 /2014/05/08/what-Useful Fusion Mutation 工具列 SEQanswers ProtocolarticlesSimilarstudiesCompany

實(shí)驗(yàn)設(shè)表達(dá)檢測，可變剪接，新轉(zhuǎn)錄物，等 表達(dá)差異，突變測，融 技術(shù)重復(fù)：基本不生物重復(fù)：樣品數(shù) Someexample Coverage(referstomappedsequencing/denovoassembly/variation

10x-30x- 30-50-100millionreadsformammalian 0.5-1million100xor20-30millionreadsper

(dependson ent,signaltonoiseandBisulfite ~30xoftheGENOMEperbiologicalSequencingstudyof$NumberfManyNumberfareproducefew

FewmoleculesareveryabundantmostreadsareattributedtoNumberofNumberofERCC ExternalRNAControls 素Assessthelowerlimitofdetectionandrange(doseAssessthefold-changeGenebodycoverage Saturation Whatinformationcanyougetfrom 功能研 部分調(diào)機(jī)制反映在RNA水平 RNA RNA 發(fā)現(xiàn)新的轉(zhuǎn) 發(fā)現(xiàn)新的細(xì)胞 singlecellRNA-LuChenetal.,Science,Whatinformationcanyougetfrom 功能研 部分調(diào)機(jī)制反映在RNA水平 RNA RNA 發(fā)現(xiàn)新的轉(zhuǎn) 發(fā)現(xiàn)新的細(xì)胞 singlecellRNA- VenablesJPetal.,NatStructMolBiolOlteanSandBatesDO,OncogeneEmergingRolesofRNAModification:LeeM.etal.,CellRNAWhatinformationcanyougetfrom 功能研 部分調(diào)機(jī)制反映在RNA水平 RNA RNA 發(fā)現(xiàn)新的轉(zhuǎn) 發(fā)現(xiàn)新的細(xì)胞 singlecellRNA-LongnoncodingCabiliNM.GenomeRes.ThelandscapeoflongnoncodingRNAsinthehumantranscriptomeIyerMatthewetal.,NatGenet.GuoJUetal.,GenomeBiol.Rybak-WolfAGuoJUetal.,GenomeBiol.Whatinformationcanyougetfrom 功能研 部分調(diào)機(jī)制反映在RNA水平 RNA RNA 發(fā)現(xiàn)新的轉(zhuǎn)