圖位克隆及重組單株

假設(shè)現(xiàn)在給你的一個(gè)課題克 試驗(yàn)中控制紫花和白花 課 地址12開發(fā)分子基于DNA-DNA雜交的DNA標(biāo)RFLP、基于PCR技術(shù)的DNA標(biāo)基于限制性酶切和PCR的DNA標(biāo)AFLP、基于單個(gè)核苷酸多態(tài)性的DNA標(biāo)3Screeningthesegregatingpopulation（indel標(biāo)記） F2 F2+-- ++ +---++ +4++PPP rawr1 89M1 0101M2 1011M3 0101M4 1101M5 0101M6 0101M7 0101 01011:0:5 whengenome/linkagemapavailable選用遺傳作圖軟件完MAPMAKER/EXP軟 Whereisthegenecontrollingfloweringcolorinpea?7Whereisthegenecontrollingfloweringcolorinpea?LinkagegroupM17,M26,M45,M56,M5,M6,M7,M76,M21,M91,

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Whereisthephenotypic (紫 2（白 （紫001（白100（紫110（白110（紫011Whereisthephenotypic (紫 2（白 （紫001紫（白100白（紫110紫（白110白（紫011紫Whereisthephenotypic (紫2（白

M7 （紫 0 （白1000（紫1101（白1100（紫0111 Whereisthe (紫2（白10101010（紫（紫00 （白10 （紫1110（白1100（紫0111Whereisthephenotypic (紫2（白

M7 （紫（紫001（白100（紫110（紫110（白110（紫011101111ConfirmtheorderofmarkersinalinkagegroupLinkagegroupOrder1:M17,M26,M45,M56,M5,M6,M7,M76,M21,M91,Order2:M17,M26,M45,M56,M6,M5,M7,M76,M21,M91,

TheorderofM5,M6, (紫2（白10101010（紫（紫00 （白10 （紫1110（白1100（紫1011TheorderofM5,M6,M7,11111200000011011000111Whereisthegenecontrollingfloweringcolorinpea? TargetCo-segregatingMarkermayBefarawayfromPurpleFlowerGene(dependsonpopulationsize) yzebothco-segregatingandflanking Inolddays:map-based

M MChangePhysical MM MPhysical Isitpossiblethatyourtargetgeneisinthecloneasamarkerthatdoesnotco-segregatewithWhydoweneedalargesegregatingTonarrowdownyourtarget FineMapwhengenome/linkagemapavailable Increasepopulationsize, 分組混合分析法（Bulkedsegregantysis,+DNA/RNA-seq(二代基于性狀表現(xiàn)型的BSA根據(jù)目將或株系依據(jù)相對(duì)表型差異分成兩組，形成座所在的區(qū)域存在差異外，來自其他部分的DNA組成完因庫的一個(gè)隨BSA+RNA-SEQorDNA-CompleteSequencingof100BC1P1 --+++--+--+++-+++-- ……---++-++-+--- PoolingExtremeP1 ++++++++++++++++ … -------------------基于標(biāo)記型的BSA 依 型A1B1/A1B1和A2B2/A2B2構(gòu)建DNA混合池，它們是在目標(biāo)區(qū)段上存在差異，目標(biāo)區(qū)段以外的遺傳尋找兩DN混合之的型有在標(biāo)上與目標(biāo) 密鎖子記。PrincipleofBSA-RNA-ATATTTATTGWhiteGCCGATTGPurpelGTTTATTTGCCTACCGATTTATCGGTCGGCTTPaperstoTraditionalmarkers(anyreviewpaper,orMichelmoreRW,ParanI,KesseliRV.1991.Identificationofmarkerslinkedtodisease-genesbybulkedsegregantysis:arapidmethodtodetectmarkersinspecificgenomicregionsbyusingsegregatingpopulations.ProceedingsoftheNationalAcademyofSciences,USA88,9828–9832.BairdNA,etal.(2008)Ra