Next Generation Sequencing

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1、ZhaoJingZhaojing@genomics.org.cn11-03-24OUTLINESequencingtechnologyoverviewCharacteristicsofthenewsequencingtechnologySolexadataanalysisThird-GenerationSequencingTechnologyGrowthofGenBankDatageneratedby1000GenomeProjecthadsurpassedtotaldatadepositedinNCBIFor

2、thesequencingcostdramaticallydecreasedsequencingtechnologyiswidlyusedinreseachCurrentChallenge:Dataminninganddisclosebiologicalissue(GeneBank-NCBI,2009)SequencingTechnologyDevelopment1950196019701980199020002010InventionofAppliedBiosystemsSolidSystemDevelopme

3、ntDiscoveryInventionof(2007)ofSangerofDNACapillarySequencingInventionofstructureSequencer(1977)Illumina(1953)(1996)GenomeInventionofInventionofAnalyzerAutomated454GS20SystemFluorescentSequencer(2006)Sequencer(2005)(1987)OUTLINESequencingtechnologyoverviewChara

4、cteristicsofthenewsequencingtechnologySolexadataanalysisThird-GenerationSequencingTechnologyNextGenerationSequencingTechnologyIllumina/HiSeq2000AppliedBiosystems/SOLIDSequencing-By-SynthesisSequencing-By-LigationRoche/454GenomeSequencerFLXSequencing-By-Synthes

5、isIllumina/HiSeq2000HIGHESTOUTPUTInitiallycapableofupto200GbperrunFASTESTDATARATE~25Gb/day7-8daysfor2x100bpHIGHESTNUMBEROFREADSOnebillionsingle-endreads*Twobillionpaired-endreads*Sequencing-By-SynthesisIlluminaSolexaFlowcellAflowcellcontainseightlanesLane1Lane

6、8•Eachtileisimagedfourtimespercycle–oneimageperbaseImagefrom1tileSimplestSequencingWorkflowHiSeq2000SIMPLIFIEDcBotCLUSTERDATAPROCESSINGSEQUENCINGSAMPLEPREPGENERATION&ANALYSISParallelsampleAutomatedclusterAutomatedSimple,efficientdataprocessinggenerationsequenc

7、inganalysisSequencingOverview/SolexaSequencingOverview/SolexaSequencingOverview/SolexaSequencingOverview/SolexaSequencingOverview/SolexaSBS（sequencing-by-synthesis）SequencingOverview/SolexaSequencingOverview/SolexaSequencingOverview/SolexaSOLiDSCALABLEOUTPUTPr

8、ovides100Gb&2000MreadsperrunUNIFORMCOVERAGEimprovecoveragetodiscoverrarevariantsindifficult(GC/AT-rich)regionsHIGHACCURACYDetectionofcausativevariationenabledatlowcoverageandcostpe