简介：Fungalpathogenscausingpoplarcankerdiseasesarecosmopolitaninspecies,andhaveawiderangeofhosts.Thesepathogenshavediverseanamorphsandtheirmorphologyoverlapswitheachother;Theirteleomorphsarehardlydiscoveredinnature.Furthermore,theidentificationofthesepathogensisusuallylimitedbythegeography,hostandtaxonomicknowledge.Therefore,aculture-independentmethodusedtoidentifydeterminepathogensisexpectedtobedevelopedforfielddiagnostics.Throughamplifying,sequencingandanalyzingmultiplexnucleicacidtemplatesandgeneticmarkedtargetsequences,amultiplexPCRtechniquehasbeenestablishedandusedtodirectlydetectvariousenvironmentalsamples.Inthisstudy,fourpathogenstrainsandenvironmentalsampleswereamplifiedusingfungaluniversalprimersITS1/ITS4andEF1-728F/EF1-986RbygeneralPCRandmultiplexPCR.Theampliconsweresequenced,andthenalignedinGenBank.TheresultshowedthatthemultiplexPCRwasabletosuccessfullyamplifythetargetgeneandidentifythepathogensfromenvironmentalsamplesintheconditionofanannealingtemperatureof55.6℃andprimersfinalconcentrationof0.2μmol·L-1.ThemultiplexPCRcouldamplifythetargetattheconcentrationlevelofapproximatelylngofgenomicDNA.ThismethodwouldprovideausefultoolfortheidentificationofcankerpathogensbythemultiplexPCRandthehigh-throughputDNAmicroarraydetectionofenvironmentalsamples.

简介：一个直角的图案被用来作为模板优化SSR-PCR扩大系统usingPanax人参genomicDNA。五个因素(DNA模板，TaqDNApolymerase，Mg~(2+)，教材，和dNTP)和退火的温度的四个层次在thissystem独立被测试了。结果证明反应效率被这些因素影响。放大aginsengSSR地点的基于的在结果，一个马厩，生产、可再现的PCR系统和骑车的节目被获得：包含1.0UTaqDNA聚合酶的20μL系统，2.0mmol·L~(-1)Mg~(2+),0.2mmol·L~(-1)dNTPs，0.3μmol·L~(-1)SSR教材，60ng·μL~(-1)DNA模板，用94的一个程序表现了为5的℃min，为30s的94℃，为30s在56.3℃退火，为1min的72℃，37个周期，为7min在72℃完成，并且在4℃存储。

简介：AllfactorsaffectingthePCRsysteminPinusmassonianaLambwereinvestigatedonebyone.TheresultsshowthattheoptimumPCRsystemofEST-SSRwere:2μL10×Buffer(Mg2+Free),30ngtemplateDNA,0.1875mmol/LdNTPs,3.75mmol/LMg2+,8pmolprimerpair,1.0UTaqDNApoly-meraseintotal20μLreactionsystem.Finally,atotalof11isolatesofP.massonianawasusedfortestingthestabilityofthePCRamplification.TheresultsprovethattheoptimumPCRsystemwasstable,reliable,highlyrepetitive.

简介：对已在橡树岭国家实验室建立起来的美洲山杨(PopulustremuloidesMichx.)和毛果杨(PopulustrichocarpaTorr.&Gray)18个微卫星引物进行筛选,用于胡杨相关基因的扩增。发现有13个基因位点可以表达基因多态性,范围是2-17个等位基因。选择了8个最易变的基因位点,来构建和优化2种多样PCR法。用来自3个种群的436株树木表征选择的位点,确定他们在亲缘关系分析、基因型研究的适用性。通过克隆基因型鉴定树木性别的交叉检验,我们估计并入基因型最大误差率小于0.045。