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    实验动物微卫星DNA、斑马鱼SNP遗传标记的检测方法、MHC单倍型实验鸡和实验鸭直接测序检测方法.docx

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    实验动物微卫星DNA、斑马鱼SNP遗传标记的检测方法、MHC单倍型实验鸡和实验鸭直接测序检测方法.docx

    1、规范性)实验动物微卫星DNA标记遗传桧测方法基因组DNA的提取基因组DNA提取方法按照NVT1673的规定执行.微卫星位点用于检测封用群和近交系实脸小型猪遗传质量的我卫星位点分别为25个和10个,各微卫星位点的名称.染色体位置、引物序列、等位基因数、等位基因范用和退火温度按照表AI的规定执行.实翳小型猪微卫星位点的引物序列退火温度、等位基因故及等位基因分布楚困位点引物序列5-3)所在於色体M广浓度(mM退火温改(C)等位签因数等位基因范H1.适用群体SW971(XiTG)UGinrrGcnniAiUi:GAAAGAMTCCAAATCeAAMIC12.05817129-175Soo91代TAC

    2、r(TA;AGATAAgccagatCGGACTCC7GCCiVJTTG21.5551496-171SW2I0AGAMTTAGTariCAAATTa;AAACCArTAAGTCCCTAGCRM21.5581192-1M封闭据SW1.O6JGMX;ATGAMXAaKrGCTCrTGAGGCAAchgctg32.06019IM2M封簿群SWIQHgr7TCCCCCACCACXJCGATCAAMTcCCTtactccgg41.55810:U-1!X)封闭群S(XM)5IaT1.XJCXnCCJGGTAKTXGCAECCTGAnCTGGGTA52.0541120i-24I封闭群SKIO57TCCCCT

    3、GnGTACAGArrGATGT(XMTKUAGTTCGgGC62.05814112-191封闭群*32TGGGnGAAAGATnCcCNXJAGTGV11RCT11GGC72.0519148173BRJiItgGCAjUtriAIICACGAAAAMNCAACCCAeTT:TCCCAC82.05912138-170封同窗SW29MiGGI1.iGC1.V1.AAMGAAMiGA7CATCCTACCGCiGC82.06112133-187封闭群SK9UCTCAGTTCmVGGACrGAMTCATCTGnXJiUAAftMMGCC92.060I1.778Bff1.etSWSUAAGCAGGAAT

    4、oCCTGCInCOCCAGCCCCAGTG,U?91.56212161-196SWr1.58TCCiVvnTAACTCCrGGCTC)02.06018158-200GAATGTGCACATAoCAaTGCSW951.11TCACAAGCA(rAGGATCGTGarAAARxK101.558M1.142SW27111CCAGT(XC11TCTGTGCCATrCArCCXAGKiAA,CrK;111.55813I1.1.-I1.1.S0386KCTGGGTCTTA1.TnCTA门TrrATCTcCAACAGrATI1.2.0481215S-17S0068CCTrGueCmGAGCMGAACAGT

    5、GGTC1.口C1.axTbTGCT132.06210210-256SWr1.oo8KACmOCAACMIGEGGAACnCcArAIGaGCAAGM132.06216!-256封闭德SOoO?11AC11C11GCATCATGTCGicccrcamAatttctg142.054IS142-192封闭辩SW85?KiAGAGGTCurnACAGAAGACCGAICCTCCgAATCCCATH2.05816129173封闭措SIfr312A1.CCGTGCGrG1.GrcCATCiGGNiGeRCAG门CUGAT151.5W111161况封闭群沏UatctggtcctxcacamjgGGGGeT

    6、sCAGGAAGGAG161.5608128-IHS*,r1.1.20CAAMGAACOCA门ACAGrOCAeTCrTNiaXAGGAGaTC171.560UM7178S(XI62)UGTCA11TAGTCAAOGTCACAGISGATMXiGAAeATAGGArAAZ182.05612IM-201封闭群SO21KGTGTAGGeTGaGGTTGT(WGRMCCIAAAGCMAGX1.55411CGAJVIAGAGmTGTAAGnGC丽GAACECArAIWCTAAGGmIq2.55512250-320近交系SW210AGAAATTAGr(XrKAAATrGGAMCrAmwiaXTAGCAA

    7、A2P1.55586-115近交系S172ATaGAKAGH;C(XXGTGTnGAAAATGGGGIGECC3P1.555HKX)-116近交系S(XIO5TtcrrcCCTCCTCGTAACrAaAcnEGATTC1.Gam5q3.055IO206-24H近交系S!WCCMGACNiOniGrAGGTGAATAGCTOCMGTATTGTAOCRrrAGC12q1.5551244-251近交系SK769GGTAIGA(XAAAMICCrcGGTsGCrXTGnXiGAAGMTGC133.05571.()6-MO近交系SW857WAGA(石Ie1.GITKAGAAGACCGatcctcxttc

    8、camtcocat141.5556IM760近交系SO355TCT(WcmCAnCbTCnGA监GTTTGGGTWnGCTGAAAMTAGCA153.05514213-277近交系!2ICnTCGGniGAGT(HGTGC八TCCAAATGeTGCAAGOG171.55586-121近交系SO218GTCrAGGCTGGaxJI1.GTCGArAAAGCMAGX1.555KIM-184近交系用于检测实粉械猴遗传质盘的微史星位点为20个,各做卫星位点的名称、引物序列、等位基因数、等位基因范用和退火温度按照表A.2的规定执行.实验荻猴微卫星位点的引物序列、退火温度、等位基因数及等位慕因分布范围位点

    9、引物序列(5-3)返火温质(C)故大等位茶闪散售位葩因分布范惘CAJA1.GAAGAOGGGGGOGTiUATATGTGGrcGCTGVTACCTGAA609386102CAJA6GAGCAOCAAGAnGGCAmCCAArMACNOGGCrnGA606235-243CAJA1.OAcoeTKATJGoCAAATrkGCCTcnCrcAGGGAAGrGA606198-20CAJA1.1.Cgj1.ugigtgctcaacaggaAMGr(XGAneIgaaagca605254262CAJA13AGCMAIGAACAaTAGGnAGIGUAKAGGCTGamGA606378-390CAJAHAG

    10、CMA1.GAACKCCAGGrTAG1.GAAKMJGCTG;AGA606216232CAJA17GGGCACTCCAAGCTCAGmTTGeC(XrrGCrrArICTAG6010382400CAJA18NTTGCAGaXagtgttctiTGGACAGCTCAGGTrrccn638315329D10qha115ICGaJMneAAAGGCGTfeTGGAGGATrcGCA11GGG606103-117Iian1.STCAGCCACTOCTCTCAGIiG1.KiGAATMArCAatrACCAG606193-205Hanj60TGCTCAGAGG11CCACTCTG(JGCATGnACC

    11、TAACCTCIV11;5881391.aHan65TGAGAACGACTGCrCrAGGTTGCAAGTCGCTrcAmCCKi5810177-201Han1.S1.CaaigagatgtgjccaagkjagCCAAACACcCAXTATaAGT587213-239IhmiaiGGOGCAGCrcArcrCrrCACCCIaXXA(ATCTTCAAGAr637150-172I1.an1.25G11XX7AAAf(IGCCArCGECAACreCTGCGTmGTC598187-201Haro0AGACCAAGCATCHbTGGArCAeCmAAACrGCTGT%TG596281-297HmM

    12、i1.CAAAGAmTGGGGATGGAAAGAmTGCAOGGCGT1UG6110263287Iian32GaXAAATeCIG1.TRiACAC5871H3-199CCACCAGA1CA1CGAGAG1AGIIani1.OOGAoCAACrOCAMGCTACGAGGrAACATGercKGACCrr588244-262I1.an26GCAAAnanGMGCATrOCAACAGnGGATGAGnIeCAG598182206用于检测实验长爪沙尺遗传质僦的微卫星位点为28个,各做卫星位点的名称、引物序列、等位基因数、等位基因范围和退火温度按照表A.3的规定执行。女脸长爪沙显微卫星位点的引物序列、

    13、退火温度,等位基因数及等位基因分布范围位点引物序列(53)退火湖取(r)Jft大等位底因效等位基区分布范IHAF200942CAGGCACOCAG11TG1.CrACACAGGCfGAGCArGT5415180-215AF200943XCTccG.mcrAC11CAAeCATTGOCMeTCTC57)71M-181AF20094481GCnTAATGTITAmXXCTVACC11CT5419113-134AF20094611TCTOGGGCTC11TCTCOCA11CrcCAGCTCCTC5728195-242AF200945AGTXCATACATCCAGUG11AKCGC.UAG(rA.G5

    14、712166-186AF20WIITGGG1.CCnrGGAAGArcCC11AVTGAATCACA5524115-153AF200947GACAGAG1.GOGAGGGGTArG1.rcGCA(i11TGG11G11TGA5517188-212D1.6Mt7CTGoCAaXctgmccattaCMCAAGA1.GTGGGGCATGA52.615180529D16Mit2Gmxjaataa1UjtataatggggtgcOCCA1.GA1.CGTMGGrnT49.19207-266D1.Mi1362TGG1.GACCGGUGATGCTGAGir8CrAAMTRnCCnG50.016176504

    15、D8M1118!GTrmeTCAGAXGIuTGCAATATAoCTaGAMAATGAGGAiVnTG1.cC48.1I1.1.%-229Dnh1.33TCTGAMnrGA*rcG11GTCGITreAAAATanGTCAnTn1.C47.315376-S94D6Mi1.37ARGA11GCAGVrcCACT(XIXiarA(JGATGTITW;AGG17.014246265DSMit31TtAGGGCTCTCTiUGGGACAAaAFtX.AGCCAaA1UFCC53.193183SOD12Mit201OCCTCGATGCCUCGCArGIG117CAACCACCCG53.118215-28

    16、3D2Mii22GCTOCCrnOc1.rrrGAACCxxxcrrrFcrT11tcc49.1g173792D1.5Mit1.21AggagacaaocaactgctgcGGCCAGrcATCACTrTATAATGC59.817232258DIIM:a.OeMAACn门GCrcCnCCGIGAGcccTAGG1.mGIGA58.71524-256D7Mit1.CCA(XT霜AArACATGIAMXCTMGATCCAiViAGATGajrTAAGC19.1I1.165-200D2Mit76CTCAAGTCraCirTCTGCCAKA(XeAAG(HTGACC1.vTG47.319281328I

    17、BNitJ30AACACATGAMCGTGTGeGTTGATAGGCATGCrTAAGCCCSO.6I1.213-251D19Mit1.TrTGTCCTCA7CAGGCCatgamagtccxgtmaakctc19.115133-165D1.IMiU35KrAACAIGGAACATeGAC(XWmiCTC【GGAGIXJCTA18.113287-307DIn1.it38CctcigaggmtaaccaagccCACAGAG11CTACCCC(XC52.6b1.1S5-2SIDXMit1.7CCTGTTMGGCACCrAGATAATAACCaVTGETCTGTGGGIK.192IM=2S1.Dt

    18、tHt36KXCHiAGAGACaVrGAGrKA(XGAGrTCKTKCCKMGTcTcCSO.69100-126DIs1.it66IEXTMGImCACAMCGacaitccttaagagagacagtoc51.7b1.272-298D13MidTCATrcAACAncrcTCAATCGCACAACAAGGnMCCTCTaGACA19.014104-132用于检测封闭群实验雪招遗传筋量的做卫星位点为21个,各微卫星位点的名称、用物序列、曳复等位基因数按照表A.4的规定执行.实骐雪貂微卫星位点的引物序列、等位叁因数及等位瓜因分布范困位点引物序列5-3)等位基因分布范恸小红等位基因数Mpu4w

    19、1.CACTTCCrCCCAIGGACACTCAAAGTCTCACCCATGC152-174(AC)23MpUA1.OvtTGGCCTArATGTGCAGATGACTCTTTGTCnGTAcCcIcTGAcCIM-158(AC)13(AC)7MA121,OGOCArCAGG1.CAICTAGGGGGTAGACACCTCGcTCAAG105-129(CA)11MpuA129GGCCTC7GACACTAG11GAAGTKAGAATaMGGATCrG1.*205(CA)13WjxiA212CCC1.A1.GAGGGCArG1.TTGTCrGCaTGnTOCACTOGT137-153(TG)13Mm1

    20、A223GAAGKAGcACOCCAGMTCTGGnGCCAAGMaAGCAG213-239(GT)19MpA229GGGTMGXCGTGCrTAAAGATGCCTCAAAGOcTeneTC107739(GT)11(TG)7WpuA231C11CCGTAACCATCTG117GTCnCAAGATGnCAGTGTGGA182-224(GT)15MptiB1.MTOCACTAeeTG(XTTCAnCAeaCWGCrCeMHCTCAG178-190(Cr)15(0)8(AC)5(AC)11MPU*2TGGGTGAGGCATGT11GCTXCAT(X(XACCirA156-164(TC)18MPU的

    21、行CCnKCAACTCTGGaGTGTGCCGGGCtG,TGA11A180-192(GA)18MpuB3AGTCGAdTCAGTGCACGAGIGxKACACATGGGVXATa175779(AG)IKAC)8(AC)7MimjK112v5CCATrACAAGfGCTKIGAGACATX;A.V:ATX:rXnVUTIXiT161165(AG)14MpuB202w3TcTCcKCmCCraTrcA1.GGATTGACaMXJA1.CAIM-168(C)5(HMpuK2O9r2crccocrGAccrCiCCCAA(JTirCCMT119-125(CT)16MpuR217w3TrcCcTCCT

    22、TCTGCTcrcnTgGG1.AM;GG1.Att汀AMC126-141(TC)S(TC)17MpuC4w7CI(iGCXrcrATCACAACATA11CGGAAGTATKTCAHXCrGCMIS763(TCCA)9MpuC1.02*3GGGTGGATGGGrcAGTAGGTAC11T0CCACTCC7C11T119-135(TGGA)7MpuD207v5(jVi(11XJA(7MpuD231v2EgggttcicacactaggtgATGCC7CAAT0CTGCTCA132-160(GATA)9用于检测实验猫遗传质地的做卫星位点为37个,石做卫星位点的名林,染色体位脏、引物用列退火温度按

    23、照表A5的规定执行.实脸猫微卫星位点的犷增条件和染色体分布位点引物庠列(5-3)染色体MIJ”浓哎(ni1.)道火湖收(IC)FCz23(TtcacactgcgagaggactTCTGACAGaTOCMGTTAerA1.1.557FCAIo62(TTCKCACGCGGGGACTTCTGKAGaToCXGGTTACrA1.1.558FCA522GrnGM11OGGC11OCCMCTTCTGGAGMCMG1.aGAAAAGIB】1.554FCA275TcactcctggacttmccatcagB21.557TGaA1.TGACGA.MXXGCAGFCA848CKC11CACAAeMXX1.GGAT

    24、GGACAI1.CrcGA.UTCCGGAGB31.557FC6M,WiAA1.UATGACTACXXACATGGTrGAGTGCGGCrM7ZCTa;GC1.1.S56FCA346,A1GOCTAGGGCCTCTCCCoCAgtcacccicctg门GC21.558FeA920CfAGrTGUGG(XecAGCACTCIGTCCAXCCTAGGGTD1.1.559FC221TGGCOCArrAAGC1.UGAAGGrTXICAT(XXA(Xr(1131.557FCA1.O1.tCrcGCAAGAAGTCCAaGGGCATGGCTCUTGGGKKWTE31.558FC1015CTtCAGTGa

    25、CAZtOGTniTT(XICCC1.UGIGirT11.555FCA1240TCtTGATGKiGCAGnAAACCGCTOCCTGACC11TCT1.)21.55FCAI76GGAAMTRiGMAGCAAAACCTeCACAGrTGGAGrTcTTAAGGA1.1.55FeMM5TGAAGA*AGMHacgcwtgGTATGAGCATCKTGTGncaTG111.55KA723TGAAGGCTAAaciCGATAGCGCAMGRTA(XGGGGGTAA1.1.55KA0R5GGTCCICWGITrT(XTATGTCTGrATGAGATGCGGTE21.558FCAO附TAGGTGANGrr

    26、cGGATnATCGMCTGAAGMC11G11GGA1.1.555FCA2IOTGAGcCAccrMiaACTC仃,ViAAGCATOCAGIGACMTGGKi1.559KAI53M1.inGAGAACUGCrCAGOGATCCTCTAN;GCAGGA(TnGA1.1.560KA672AAGTTGcrrccAGocrccTCXAM;AGrmTCAG1IMX;E21.560FWD56GGTGMAGGGCmTTCrcAGGTTCTCCCTTGCTGGT1.55KA1239AAAAaXCnIacacccaagcrrGccrAv11GcrcA11;1)21.558FCA700CcCTIMVAT。;G

    27、IGcrC1.GMtccaawmaacaggcctB1.1.55FCA742TcAArGTCnGACAACGCATAA,GA11GCTGCCAGGMCD1.1.556K678TCarrCAGCAIUVnr&;AAGMGGAGEGCTGAAAnGrrAI1.556KAO96CACGCCAACICNWCMACAATGTGaiGTOCNAGAACE21.55FCAO75AIGC1.AATCAGNiGCAmGGGMciUAAATrCCAGACGIGC121.558FCA1I9CCArc.Wi11OCCCAAAFCGKTCACCATCTGHXiGATGB1.1.55FCA220CGATGGAMTTGN1

    28、CCAIGGGaatgaaoxmtckaaactg1.558FCA229CAAACIGMAMicnAGAG(JGCGCAGAAGK1.CHTCrCAAAGTCA1.1.558用于检测实验,鸣遗传质豉的微卫星位点为29个.各做卫星位点的名称、引物序列、等位基因数和等位基因范围按照表A.6的规定执行,实物鸿微卫星位点的引物序列、退火温度、等位基因数及等位思因分布范困位力引的序列5-3)退火粗度(TC)大等等基因数等位基因分布更BIMCYOO29CrcGACAOeCAnTGTAarrA1.GCATCCAArrcAGGAOeCTGcA6X8139188AD1.O293CrAATaGAcc0?MaCT

    29、CCQCAGKrGC5X96HXi-120AD1.o317G1T(XT11CGCCA7CCATOXMA(JCAScTGTCAUGS8.5177-219GaoDI6CCAAGGrcTrAGTTCGGCRTAAoGAT八GCAACAG52.26111-118AD1.OIMtI欢GAGGAACIrTGGAM1.GCCTanGCncGTGCTETSX97KW-1611.KI0074GACCmGCCGMWIWTGGrnGerGATTAGeCAToGCG58.56221-243AD1.328CAOTATAGSGTGACrnG53.95107-120AAAAcCGGAArG7GTACTGGGANrECIGO

    30、GG(XCOGTATCGAGCARGgGGGCGCrCCTGGT65.07139-1941.EI0094*CAGGATGGaGmTGCrTccACACAGTGeAGAgtggigcga56.07176211MCMoa30TGGAOCIRTCAGTCTCACAGAMGnCrCArAGMTrOcTCC58.562172871.EIOH1.CGCmGAgrAACACAR;MGGeMKTeAGCNiGAAeG52.25221245MCMo1.7ATneIGQGCcUC1.TGGAGC1.eUX;CAGTnJCAGAC58.5g268289MC0i47GCnOCAGATGAGCrcmGGCAC&X:GC

    31、M1.AGCAACwS2.03M9AD1.176TCT(XA11CT(X(XAGC门ETarCGrAKAC1.QnCA58.59183200AW.0201GCTGAGG17CAGAT.UGACMgTGACGnTCACAGC58.57111-151G(MAMZOGTCACTAGGTrAGCAGCA1.GGCr(XiATACAG)UritGAFT56.0123AGGAYIRGAGA1CGIGCACGCU(11CGAGCACnTCNiGCAGAGA60.73S6-K9M1.YoO63GGerOCAAAAGCrTGTrCnKCrGAAAA(XMmuAGaTCbKSX98116-14AD1.1.85CA

    32、1GGCAOC7GACCCGATAGCGnAOCTGnCGTrrcCSB.S9116-142CXJMYCCGAG(XGcKr(XGAG1.TIATGGGGACercrGGCrcTGAC62.45B9-151.1.KI009tGA1.CKACCAGrAfGAGCIGCTCTCKReTGTAAeKMTGCS3.96250-283GGY1.rC(XrCA7CV11CA(iGOKTGATCKrniACTGATCCrcAS8.52S690AD1.O292CCAAA1.CAGGCAAAACTTaATCGCCTAAGG*TGAGGA58.56110-136(XJV1.TIJG*(NHMGTnrrAATarW

    33、ACeCATCGECAACimTGS6.02ADIJ66rGCCAGCOCGTAATCATAGGMGCKCKGReCCAATCTA58.56131-154MCV1.OO1.tAAAATATTCGatTAGGAACKSTCACCGGAAATGAftGGTAAGACTAGCS8.59172-195MiCGAGGOGCrcrGtGGIIGGGCACCTCTGGCGACC61.6510-153MCYO1。2CI6M(T1.MXiAC1.AGaGCcTAAGTCTGGGCTenTTGS6.015111-229WNSGGH1.:2TACnAATArGTGrcAGGT(XCGnCItAeAArrcCATTA

    34、GC5X92235238用于检测实粉料遗传质成的微卫星位点为29个,各做犯星位点的名称、引物序列、等位基因数和等位基因范用按照表A7的规定执行.实蕤鸭微卫星位点的引物序列、退火温度、等位基因数及等位基因分布范困位点引物序列(5-3)退火温慢(C)最大等位基闪数等位基闪分布范困AP1.2GAncAACCTTAGCTAICAGTCTCCCCCTtTTGGCAMTGKr58.54115-125AP1.579A1.TAGAGCAGGMTTAGGKACGCAAGAAGTGGCnTnTC557118-227APHo9GGA1G11GOCCCCTA1111GCC11G11ATGAGCCA1.TA588937

    35、88APH1.1.GGACCTCAGGA1.UACGTGTGCAGGCAGAGCMGAA58.52IM*185RPHI8TrcTCGarGArAGGfATGAG(hUrTXiGUiGItATACIXiT583179-324AY258ArcicrGAGICCT1.CGG.MXACAATACArrCCAGaTGCTGAA58.195)761AY264GCMACnnAC1.TA愉CTCCnAGeCCAGTGAAGCATG58.111U2328AY311CTCATnXM1Ta:Te1.CfACGCrA11T11CAGAAGG50.33IS5-250CAVD1ACAG(TTeMGgUTMGAGCAGAA

    36、AG1.1.GTaTrAAGGAAG55.522M-390CACDOO2CnaxnGOcgImAGCAGCHxxIGGAGAAGG60.i175-231CAtDOOI1CCC11GGJAGACC11GAGGGGArrCAGGAGAAGCa60.83162-290CA1.D005CT(XGmanGGAGCArAArcr(xxriGccr11r11;60.3180-30()CAmOa6AnxTTcreTGrAGGCAMTTrCKiCIIGGGC1.XTNiGA63.58I3-21CM:DanACncrcrrGTAGGCA1.GTrACa(X1.GnGCTCerGaGT60.86KW-2OHCAUD

    37、010GGATCrGTErcArrAr1.ETAgmgcatmatktcagtg50.39IKO-3O()CA1.IDO)1IGCrAKCKCCAATAAGWCG11GCTGGTATCTa:50.36I:47-222CAtDO12ArrcCCTnCAGrcGAGnTCCGGCrCTAMCRCATGAATG6X59182-2H6CAt!D011.C1.CAAeIGKGGCAcAAMT58.13136200CrGAGrn门CaXCCTCsCAID026CG7CCATCACCACAGCmCCCrC1.CGTGKG门T60.89U196CAUDO27AGAAGGCAOGCAAATCACKIirACiCATAAnaacacccaca66S100-180CA1.IDa28IACACOCAAGTnArKTGAGACTerOCAoGGCACTAGG55.5X153-220CAUDa31MCAIeTGGACITr口Cr(XACCCCCAGGC1.CTgGATAA51.410MO-187CAUDa32GUAeCAAcIGAAAWG(XC(TmKEaxmTAN58.19邻一206CAUD(XM7AGCATA1.CACTAG,GGAT(XCATACTCGGG11AC55.59160-2%CAID035GnXCrAAOCCrGAnXATGCHATCAGAIGaiGaCGGA63.5


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