當前位置 ： NEB >>> NEB/Luna? Universal Probe One-Step RT-qPCR Kit/E3006S/1,000 reactions
NEB/Luna? Universal Probe One-Step RT-qPCR Kit/E3006S/1,000 reactions
  • NEB/Luna? Universal Probe One-Step RT-qPCR Kit/E3006S/1,000 reactions

NEB/Luna? Universal Probe One-Step RT-qPCR Kit/E3006S/1,000 reactions

價格: ￥9996.00 市場價: 16660.00
品牌: NEB
規(guī)格
數(shù)量
庫存(0)
特別 提示
代購產(chǎn)品：無質(zhì)量問題不接受退換貨，下單前請仔細核對信息。
下單后請及時聯(lián)系客服核對商品價格，訂單生效后再付款。
資深產(chǎn)品顧問
咨詢顧問

全國免費服務(wù)熱線

4000-520-616

  • 自營商城 一站式服務(wù)
  • 廠家直采 剔除溢價
  • 品質(zhì)甄選 正品保證
  • 嚴控流程 只做188精品
  • 極速物流 如約送貨
  • 詳情
  • 使用說明
  • 常見問題

    • Description:

      Sample
      ?

      Rapid,sensitiveandpreciseprobe-basedqPCRdetectionandquantitationofRNAtargets.

      TheNEBLunaUniversalProbeOne-StepRT-qPCRKitisoptimizedforreal-timequantitationoftargetRNAsequencesusinghydrolysisprobes.One-StepRT-qPCRprovidesaconvenientandpowerfulmethodforRNAdetectionandquantitation.Inasingletube,RNAisfirstconvertedtoCDNAbyareversetranscriptase,andthenaDNA-dependentDNApolymeraseamplifiesthecDNA,enablingquantitationviaqPCR.Probe-basedqPCR/RT-qPCRmonitorsanincreaseinfluorescenceupon5′→3′exonucleasecleavageofaquenched,target-specificprobetomeasureDNAamplificationateachcycleofaPCR.Atapointwherethefluorescencesignalisconfidentlydetectedoverthebackgroundfluorescence,aquantificationcycleorCqvaluecanbedetermined.Cqvaluescanbeusedtoevaluaterelativetargetabundancebetweentwoormoresamples,ortocalculateabsolutetargetquantitiesinreferencetoanappropriatestandardcurvederivedfromaseriesofknowndilutions.

      IntheLunaUniversalOne-StepProbeRT-qPCRKit,HotStartTaqDNAPolymeraseiscombinedwithanovelWarmStart-activatedreversetranscriptase,allowingdualcontrolofenzymeactivityviareversIBLe,aptamer-basedinhibition.Thistemperature-dependentactivationhelpstopreventundesirablenon-specificprimingandextensionpriortoThermocycling,providingaddedsecurityforsettingupreactionsatroomtemperature.TheengineeredWarmStartLunaReverseTranscriptasealsopossesseshigherthermostABIlitythanmanyotherRTs,allowinganoptimalreactiontemperatureof55°C.Fordifficulttargets/templates,higherRTsteptemperaturesofupto60°CcanbeusedwithoutcompromisingLunaperformance.
      ?
      NotethattoensurefullactivationoftheWarmStartLunaRT,incubationattemperatureslowerthan50°Cisnotrecommended.

      TheLunaUniversalProbeOne-StepReactionMixissuppliedat2XconcentrationandcontainsHot-StartTaqDNAPolymerase,dNTPs,andallrequiredbuffercomponents.Itisformulatedwithauniquepassivereferencedyethatiscompatibleacrossavarietyofinstrumentplatforms,includingthosethatrequireahighorlowROXreferencesignal.TheReactionMixalsofeaturesdUTPforcarryoverpreventionandanon-fluorescentvisibledyeformonitoringreactionsetup.ThisvisibledyedoesnotoverlapspectrallywithfluorophorescommonlyusedinqPCRanddoesnotinterferewithreal-timedetection.

      TheLunaWarmStartRTEnzymeMixissuppliedat20XconcentrationandcontainsLunaWarmStartReverseTranscriptaseaswellasMurineRNaseInhibitortoaidinpreventingRNAdegradation(seealsotemplatepreparationinproductmanual).ItiscompatiblewithvariousRNAsampletypes(totalRNA,poly(A)-RNA,etc.)andsources.

      .

      Figure1:NEB’sLunaUniversalProbeOne-StepRT-qPCRKitoffersexceptionalsensitivity,reproducibilityandRT-qPCRperformance
      RT-qPCRtargetinghumanGAPDHwasperformedusingtheLunaUniversalProbeOne-StepRT-qPCRKitoveran8-lograngeofinputtemplateconcentrations(1μg–0.1pgJurkattotalRNA)with8replicatesateachconcentration.Reactionsetupandcyclingconditionsfollowedrecommendedprotocols,includinga10-minuteRTstepat55°CforthethermostableLunaWarmStartReverseTranscriptase.


      Figure2:NEB’sLunaUniversalProbeOne-StepRT-qPCRKitoffersrobustperformanceinmultiplexapplications
      Luna
      MultiplexRT-qPCRtargetinghumanGAPDH,ribosomalproteinL32gandPI3-Kinase-RelatedKinaseSMG1wasperformedusingtheLunaUniversalProbeOne-StepRT-qPCRKitovera7-lograngeofinputtemplateconcentrations(1μg–1pgJurkattotalRNA)with4replicatesateachconcentration.Amplificationplotsareshownbothoverlayed(left)andforeachmultiplextarget(right).Toaccountforcopynumberdifferences,0.4μMprimerwasusedforlower-copytarget(SMG1)and0.2μMprimerforhigher-copytargets(L32gandGAPDH).Lunamaintainssuperiorefficiency,reproducibility,sensitivityandperformanceinmultiplexRT-qPCR.



      Figure3:Extensiveperformanceevaluationofcommerciallyavailableprobe-basedRT-qPCRreagentsdemonstratestherobustnessandspecificityofLuna
      Luna
      Commercially-availableRT-qPCRreagentsweretestedon7RT-qPCRtargetsvaryinginabundance,length,and%GC.Datawascollectedby2usersandaccordingtomanufacturer’srecommendations.Resultswereevaluatedforefficiency,lowinputdetectionandlackofnon-templateamplification(whereΔCq=averageCqofnon-templatecontrol–averageCqoflowestinput).Inaddition,consistency,reproducibilityandoverallcurvequalitywereassessed(QualityScore).Bargraphindicates%oftargetsthatmetacceptableperformancecriteria(indicatedbygreenboxondotplotandQualityScore>3).ResultsforNEBandothersuppliersareshown:Quanta,qScript?XLT1-StepRT-qPCRToughMix?;ABI,TaqMan?RNA-to-Ct1-StepKit;Qiagen,QuantiFast?ProbeRT-PCRKit;Bio-Rad,iTaq?UniversalProbesOne-StepKit;Promega?,GoTaq?Probe1-StepRT-qPCRSystem.NEB’sLunaUniversalProbeOne-StepRT-qPCRKitoutperformedallotherreagentstested.

      LearnmoreaboutourcomprehensiveqPCR/RT-qPCRtestingand“dotsinboxes”datavisualization.

      KitComponents

      Thefollowingreagentsaresuppliedwiththisproduct:

      Storeat(°C)Concentration
      Luna?UniversalProbeOne-StepReactionMix-202X
      Luna?WarmStart?RTEnzymeMix-2020X
      Nuclease-freeWater-20

      Notes:

      PrimerDesignTheuseofqPCRprimerdesignsoftware(e.g.,Primer3)maximizesthelikelihoodofamplificationsuccesswhileminimizingnonspecificamplificationandprimerdimers.TargetswithbalancedGC/ATcontent(40–60%)tendtoamplifymostefficiently.Wherepossible,entersufficientsequencearoundtheareaofinteresttopermitrobustprimerdesignandusesearchcriteriathatpermitcross-referenceagainstrelevantsequencedatabases(toavoidpotentialoff-targetamplification).ItisadvisabletodesignprimersacrossknownRNAsplicingsitesinordertopreventamplificationfromgenomicDNA.PrimerandProbeConcentrationsFormosttargets,afinalconcentrationof400nM(eachprimer)willprovideoptimumperformance.Ifneeded,primerconcentrationscanbeoptimizedbetween100–900nM.Probeshouldbeincludedat200nMforbestresults.Probeconcentrationcanbeoptimizedintherangeof100–500nM.MultiplexingWhendeterminingwhichfluorophorestoincludeinamultiplexreaction,besuretochoosecompatiblereporterdyesandquenchers(e.g.,thosethatcanbeaccommodatedbythechosenreal-timeinstrumentwithminimaloverlapinfluorescencespectra).ForROX-dependentinstruments,avoidROX-labeledprobes.Include400nMofforwardandreverseprimersand200nMprobeforeachtargettobedetectedinthereaction.Fortargetsthatdiffersignificantlyinabundance,useofalowerprimerconcentration(e.g.200nM)forthemoreabundanttarget(s)isrecommended.Adjustconcentrationsifnecessarybasedonperformance(primer100–900nM,probe100–500nM).WhenloADIngaqPCRprotocolontothereal-timeinstrument,besuretoselecttheappropriateopticalchannels,assomeinstrumentshaveasinglechannelrecordingmodethatwouldpreventmultiplexdatacollectionandanalysis.Thefunctionalityoftheprimerandprobesetsshouldbetestedindividuallybeforeattemptingamultiplexreaction.AmpliconLengthToensuresuccessfulandconsistentqPCRresults,itisimportanttomaximizePCRefficiency.AnimportantaspectofthisisthedesignofshortPCRamplicons(typically70–200bp).Someoptimizationmayberequiredfortargetsthatexceedthatrange.TemplatePreparationandConcentrationLunaRT-qPCRiscompatiblewithRNAsamplespreparedthroughtypicalnucleicacidpurificationmethods.PreparedRNAshouldbestoredinanEDTA-containingbuffer(e.g.,1XTE)forlong-termstability,anddilutionsshouldbefreshlypreparedforaqPCRexperimentineitherTEorwater.NotethatthequalityofRNAtemplatescangreatlyaffectRT-qPCRefficiency.RNAshouldbehandledwithappropriateprecautionstopreventRNaseorDNasecontamination.Useofnuclease-freewater(provided)isstronglyrecommended.Whereuseful,RNAmaybetreatedwithDNaseItoremovecontaminatinggenomicDNA.Generally,ausefulconcentrationofstandardandunknownmaterialwillbeintherangeof108copiesto10copies.Notethatfordilutionsinthesingle-copyrange,somesampleswillcontainmultiplecopiesandsomewillhavenone,asdefinedbythePoissondistribution.FortotalRNA,LunaOne-StepKitscanprovidelinearquantitationoveran8-orderinputrangeof1μg–0.1?pg.Formosttargets,astandardinputrangeof100ng–10pgtotalRNAisrecommended.ForpurifiedmRNA,inputof≤100ngisrecommended.Forinvitro-transcribedRNA,inputof≤109copiesisrecommended.ROXReferenceDyeSomereal-timeinstrumentsrecommendtheuseofapassivereferencedye(typicallyROX)toovercomewell-to-wellvariationsthatcouldbecausedbybubbles,smalldifferencesinvolume,andautofluorescencefromdustorparticulatesinthereaction.LunamixesareformulatedwithauniversalreferencedyethatiscompatiblewithavarietyofqPCRinstrumenttypes,includingthosethatusenopassivereferencenormalizationandthosethatusealoworhighconcentrationofpassivereferencedye(ROX).Therefore,noadditionalcomponentsarerequiredtoensurecompatibilitywiththeseinstruments.CarryoverContaminationPreventionRT-qPCRisanextremelysensitivemethod,andcontaminationinnewRT-qPCRassayswithproductsfrompreviousamplificationreactionscancauseavarietyofissues,suchasfalsepositiveresultsandadecreaseinsensitivity.Thebestwaytopreventthis“carryover”contaminationistopracticegoodlaboratoryproceduresandavoidopeningthereactionvesselpostamplification.However,toaccommodatesituationswhereadditionalanti-contaminationmeasuresaredesired,LunaqPCRmixescontainsamixtureofdUTP/dTTPthatresultsintheincorporationofdUintotheDNAproductduringamplification.PretreatmentofqPCR/RT-qPCRexperimentswithuracilDNAglycosylase(UDG)willeliminatepreviously-amplifieduracil-containingproductsbyexcisingtheuracilbasetoproduceanon-amplifiableDNAproduct.TheuseofathermolabileUDGisimportant,ascompleteinactivationoftheUDGisrequiredtopreventdestructionofnewlysynthesizedqPCRproducts.Toenablecarryoverprevention,0.025units/μlAntarcticThermolabileUDG(NEB#M0372)shouldbeaddedtothereactionmix.Tomaximizeeliminationofcontaminatingproducts,setuptheqPCRexperimentsatroomtemperatureorincludea10minuteincubationstepat25°Cbeforetheinitialdenaturationstep.ReactionSetupandCyclingConditionsDuetodualhot-startfeatureofLunaOne-StepKits,itisnotnecessarytosetupreactionsoniceorpreheatthethermocyclerpriortouse.For96-wellplates,afinalreactionvolumeof20μlisrecommended.For384-wellplates,afinalreactionvolumeof10μlisrecommended.Whenprogramminginstrumentcyclingconditions,ensureaplatereadisincludedattheendoftheextensionstep,andadenaturation(melt)curveaftercyclingiscompletetoanalyzeproductspecificity.Amplificationfor40cyclesissufficientformostapplications,butforverylowinputsamples45cyclesmaybeused.
    售后保障
    螞蟻淘生物188，秉承螞蟻淘一貫的嚴謹態(tài)度，由螞蟻淘公司專業(yè)人員負責(zé)品控、采購、物流、銷售、售后，保障正品優(yōu)質(zhì)。以“快速好省，為科研提供好產(chǎn)品、好價格”為理念，直接鏈接原廠家，從全國各地原制造商嚴格挑選188款科研精品，剔除品牌溢價，188生物新電商，把好的產(chǎn)品帶給科研！? 力求給你最優(yōu)質(zhì)的商品。
  • Q：生物188產(chǎn)品正品保障嗎？
    A：生物188質(zhì)量把控人員具有十年的從業(yè)經(jīng)驗，在業(yè)界享有良好的口碑；自營商城，直接從廠家采購， 自己的團隊負責(zé)國際物流和清關(guān)，中間沒有第三方，所有流程嚴格把控，100%保證正品，假一罰十。

    Q：下單后可以修改訂單嗎？
    A：下單后的商品付款之前可以修改；訂單付款成功，需要聯(lián)系我們客服進行修改；客服電話：4000-520-616

    Q：可以開發(fā)票嗎？
    A：本網(wǎng)站所售商品都是正規(guī)清關(guān)，均開具16%正規(guī)發(fā)票，發(fā)票金額含配送費金額，另有說明的除外。

    Q：商品幾天可以發(fā)貨？
    A：生物188商品，全部現(xiàn)貨銷售，付款后即可發(fā)貨,一般一周內(nèi)送達！

    Q：如何聯(lián)系商家？
    A：有任何問題夠可以電話咨詢我們，全國24小時免費服務(wù)熱線：4000-520-616 或聯(lián)系我們的在線客服QQ：1570468124

    Q：收到的商品少了/發(fā)錯了怎么辦？
    A：同個訂單購買多個商品可能會分為一個以上包裹發(fā)出，可能不會同時送達，建議查看訂單詳情是否是 部分發(fā)貨狀態(tài)；如未收到，可聯(lián)系在線客服或者致電4000-520-616。

    Q：退換貨/維修需要多長時間？
    A：一般情況下，退貨處理周期為客戶收到產(chǎn)品一個月內(nèi)（以快遞公司顯示簽收時間為準），包裝規(guī)格、 數(shù)量、品種不符，外觀毀損、短缺或缺陷，請在收到貨24小時內(nèi)申請退換貨；特殊商品以合同條款為準。

何為188

極簡而嚴謹，我們僅銷售188款生物醫(yī)學(xué)科研用品，款款都是爆款；因為少所以聚焦，聚焦甄選每一款產(chǎn)品，聚焦服務(wù)每一位客戶!

關(guān)注我們 :

點擊QQ聯(lián)系我們
生物188微信

關(guān)注188微信公眾號

獲取最新優(yōu)惠活動通知