AB75002.0绝对定量简易操作流程

1、appliedbiosystems7500荧光定量PCR仪绝对定量实验简易操作流程SDS2.0绝对定量实验简易操作流程口 DItechnologies7500定量PCR仪绝对定量实验简易操作流程1.双击桌面图标，或从StartA11ProgramsAppliedBiosystems7500Software7500V2.0开启软件。进入主界面后选择AdvancedSetup2.默认进入Setup下的ExperimentProperties界面2.1输入实验名称(ExperimentName)Howdoyouwanttoidenticthtsexperiment?Whichinstrumentar

2、eyouusingtoruntheexperiment?St2.：3.在实验;类型中77500(曲Wells)|Whichreagentsdoyou1wanttousetodetectthetargetsequence?Whichrampspeeddoyouwanttouseintheinstrumentrun?；】2孑5确认运行:模式式SCSS!：!?：3：T；!?：；f.V；：v圧辽FEW：一沱C：i1；3邙亍*i：i:；v3!：；：；：GJ；，：W；W：2：；:?兀沢苗U氏iil：!=Foroptimalresultswiththestandardrampspeed,AppliedBio

3、systemsrecommendsusingstandardreagentsforyourPCRreactions.VTaqManReagentsVStandard(-2hourstocompletearun)Fast十40minutestocompletearun)绝对定量实验简易操作流程口 #D绝对定量实验简易操作流程口 #D3.进入Setup下的PlateSetup界面，编辑基因（Target）及样本（Sample）：绝对定量实验简易操作流程口 Dtechnologies,jNfwEcpitftnefTlJppw*5j*eQpi睁.ijSumEaf-ainNMiiir&ninuii口ii

4、vommEjEpfiinMiiIrcmircfifLMnMn4Enrt占PmiRm:.E-XpAMitafill.hililkfJ1RunrAnl/4-ic-T”SJamilaidDuijv&Ftemg的怕TaqMaMReageriilXTUTT(?JH:绝对定量实验简易操作流程口 #D3.1在|defineTargetsandSamples界面中设置基因及样品。利用添加新的基因，并在TargetName中编辑基因名称，Reporter和Quencher中选择所标记的荧光基团及淬灭基团。对于Quencher的选择，如果是MGB探针，请选择NFQ-MGB；如果是TAMRA探针，请选择TAMRA；

5、如果是其他形式的非荧光淬灭基团（如BHQ），请选择None。绝对定量实验简易操作流程口 #D绝对定量实验简易操作流程口 DAddNswTargaiIJtSamptaNama-AddSawdTa-gcl1Saira-golDolElaraiggtCclor(MineTsngetsandSarpi&susiictiarNFCMeHSB*NFQ-UGE：NenaTaigGlNameRsgDrinr3FOHAddHimSampleCdIdi1O*finTargEtsAdd3mriSamplsljInariK奇CT曲DQlnQ宙口号也Idquarty2廿mebn計口石totMlnUkiroxlNNipl

6、ate绝对定量实验简易操作流程口 DItechnologies还可以利用其他按钮将之前已添加的基因进行保存(SaveTarget)或删除(DeleteTarget)。利用AddNewSa添加新的样本，并编辑样品名称。32在朋引goTsgE也andSamp止中进行样品板的排布。利用鼠标单选或拖曳以选择反应孔，然后勾选左侧的基因及样本，同时在Task选项中指定该反应孔的类型(S代表标准曲线数据点，U代表未知样本，N代表阴性对照)。FileEditInstrumentAnalysisToolsHe-lp绝对定量实验简易操作流程口 #DJNewExperiment-_JOpen.Save-CloseS

7、endExperimenttoInstrument.DownloadExperimentfrom绝对定量实验简易操作流程口 #D绝对定量实验简易操作流程口 #DExperimentMenuExperiment:96-WellComparativ._Type:ComparativeCt(AACt)绝对定量实验简易操作流程口 #D绝对定量实验简易操作流程口 DDefineTargetsandSamples=BiologicalGroupAssignAssignsamplefsofselectedwell(stobiologicalgroup.Quantitye:GraphType:-Log厂ISa

8、velyjrenjLUlt噬IX:mfi:inosasthedef:-=1力HabuRunMethodExperimentProp.ExperimentMenuExperiment：96-WellStandar.Type：StandardCurveMuttipiePtotsView绝对定量实验简易操作流程口ItechnoloaiesFileEditinstrumentAnalysisToolsHelp厂亦丽訂勾选以显示阖值线及基线NewExperimentt.JOpen.dSavetClose总Export.t&PrintReport.更改图形显示方式(MotSettingsAmplifica

9、tionPlotExperimentMenuJ：2JNewExperiment-Open.Save-二CloseSendExperimenttoInstrument.曇DownloadExperimentfromfAnalysisExperiment:UntitledType:StandardCurve轉ReviewthereactionvolumeandthethermalprofileforthedefaultrunmD.1J_IT1111T1I,11111111134日aIO1214161320222436302239aaCycleTargetThreshold:i7Autoo软件将ee

10、xportfnnnatandioselectfieldsioExportClickStartEsparl*inexportyourExportPropertiesICustornizeExportSampleSetupRawData1Select翻审toejcpoeTAnipirikalionDaSa占主弦讎撓石T击看越申工安THOLDRAJLThresholdingalgorithmfailed0查看Fhg详细信息._iNewExperiment-Open.肩Save-一CloseSendExperimenttoInstrument.,DownloadExperimentfroitFArly

11、导祐rmyountnfaijf0LFA1LEimsElinE目lqciithmfmilEdCTFA1LCtmlgorithrnfmilEdProcessedWei.96ManuallyOmittedWei.0FlaggedWells:BAnalysisOmittedWells:0TargetsUsed:1SamplesUse.0Flag:HIGHSDHighstandarddeviationinreplicategroupFlagDetait:TheCtstandarddeviationfortherepficategroupexceedstheflagsetting.FlagCriieria:Ctstandarddeviation0.5FlaggedWels:AHaBit；C11DHnE113F11nG113H11OpenlllflisFMier.expcfliscompleio.1TotalWells:96WellsSetU.9e单总击HIGHS右丁季hsaVe口InfunTimti