Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 508/528 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | NucleicAcidDetection RNADetection |
Related | LabelingCells FluorescenceImaging BiochemicalAssays |
1.Preparing1Xassaybuffer
Preparea1Xassaybufferbydilutingthe10Xassaybuffer(ComponentB)withsterile,distilled,nuclease-freewater.
2.PreparingStrandBrite™RNAGreenworkingsolution
PrepareStrandBrite™RNAGreenworkingsolutionbymakinga200-folddilutionoftheconcentratedDMSOsolutionin1Xassaybuffer(fromStep1).Forexample,add10µLofStrandBrite™RNAGreen(ComponentA)into2mLof1Xassaybuffer. Protecttheworkingsolutionfromlightbycoveringitwithfoilorplacingitinthedark.
Note1:Werecommendpreparingthissolutioninaplasticcontainerratherthanglass,asthedyemayadsorbtoglasssurfaces.
Note2:Forbestresults,thissolutionshouldbeusedwithinafewhoursofitspreparation.
3.PrepareserialdilutionsofRNAstandard(0to20µg/mL):
3.1 Add10μLof2mg/mLRNAstocksolution(ComponentC)to990µLof1Xassaybuffer(fromStep1)tohave20μg/mLRNAsolution,andthenperform1:2serialdilutionstogetapproximately20,10,5,2.5,1.25,0.625,0.313,and0µg/mL.
Note:UnusedRibosomalRNAStandard(ComponentC)shouldbedividedintosingleusealiquotsinnuclease-freeplasticvialsandstoredat≤-20ºC.
3.2 AddRNAstandardsandRNAcontainingtestsamplesintoa96-wellsolidblackmicroplateasdescribedinTables1and2.
Table1.LayoutofRNAstandardsandtestsamplesinasolidblack96-wellmicroplate
BL | BL | TS | TS | …. | …. |
|
|
|
|
|
|
RS1 | RS1 | …. | …. | …. | …. |
|
|
|
|
|
|
RS2 | RS2 |
|
|
|
|
|
|
|
|
|
|
RS3 | RS3 |
|
|
|
|
|
|
|
|
|
|
RS4 | RS4 |
|
|
|
|
|
|
|
|
|
|
RS5 | RS5 |
|
|
|
|
|
|
|
|
|
|
RS6 | RS6 |
|
|
|
|
|
|
|
|
|
|
RS7 | RS7 |
|
|
|
|
|
|
|
|
|
|
*Note:RS=RNAStandards;BL=BlankControl;TS=TestSamples
Table2.Reagentcompositionforeachwell
RNAStandard | BlankControl | TestSample |
Serialdilutions*100μL | 1Xassaybuffer100μL | 100μL |
*Note:AddtheseriallydilutionsofRNAstandardsfrom0.3to20µg/mLintowellsfromRS1toRS7induplicate.
4.RunRNAassay:
4.1 Add100μLofStrandBrite™RNAGreenworkingsolution(fromStep2)toeachwelloftheRNAstandard,blankcontrol,andtestsamples(seeStep3)tomakethetotalRNAassayvolumeof200µL/well.
Note:Fora384-wellplate,add25μLsampleand25μLofStrandBrite™RNAGreenworkingsolutionperwell.
4.2 Incubatethereactionatroomtemperaturefor2to5minutes,protectedfromlight.
4.3 MonitorthefluorescenceincreasewithafluorescencemicroplatereaderatEx/Em=490/540nm(cutoffat515nm).
Note:Tominimizephotobleachingeffects,keepthetimeforfluorescencemeasurementconstantforallsamples.
References&Citations | PrinterFriendlyVersion |
1. PetersonEJ,KireevD,MoonAF,MidonM,JanzenWP,PingoudA,PedersenLC,SingletonSF.(2013)InhibitorsofStreptococcuspneumoniaesurfaceendonucleaseEndAdiscoveredbyhigh-throughputscreeningusingaPicoGreenfluorescenceassay.JBiomolScreen,18,247.
2. ChenY,SonnaertM,RobertsSJ,LuytenFP,SchrootenJ.(2012)ValidationofaPicoGreen-basedDNAquantificationintegratedinanRNAextractionmethodfortwo-dimensionalandthree-dimensionalcellcultures.TissueEngPartCMethods,18,444.
3. MorenoLA,CoxKL.(2010)QuantificationofdsDNAusingtheHitachiF-7000FluorescenceSpectrophotometerandPicoGreendye.JVisExp.
4. DraganAI,Casas-FinetJR,BishopES,StrouseRJ,SchenermanMA,GeddesCD.(2010)CharacterizationofPicoGreeninteractionwithdsDNAandtheoriginofitsfluorescenceenhancementuponbinding.BiophysJ,99,3010.
5. DraganAI,BishopES,Casas-FinetJR,StrouseRJ,SchenermanMA,GeddesCD.(2010)Metal-enhancedPicoGreenfluorescence:applicationtofastandultra-sensitivepg/mlDNAquantitation.JImmunolMethods,362,95.
6. AbiodunOO,GbotoshoGO,AjaiyeobaEO,HappiCT,HoferS,WittlinS,SowunmiA,BrunR,OduolaAM.(2010)ComparisonofSYBRGreenI-,PicoGreen-,and[3H]-hypoxanthine-basedassaysforinvitroantimalarialscreeningofplantsfromNigerianethnomedicine.ParasitolRes,106,933.
7. DraganAI,BishopES,Casas-FinetJR,StrouseRJ,SchenermanMA,GeddesCD.(2010)Metal-enhancedPicoGreenfluorescence:applicationfordouble-strandedDNAquantification.AnalBiochem,396,8.
8. HoldenMJ,HaynesRJ,RabbSA,SatijaN,YangK,BlasicJR,Jr.(2009)FactorsaffectingquantificationoftotalDNAbyUVspectroscopyandPicoGreenfluorescence.JAgricFoodChem,57,7221.
9. IkedaY,IwakiriS,YoshimoriT.(2009)DevelopmentandcharacterizationofanovelhostcellDNAassayusingultra-sensitivefluorescentnucleicacidstain"PicoGreen".JPharmBiomedAnal,49,997.
10. RenX,XuQH.(2009)Label-freeDNAsequencedetectionwithenhancedsensitivityandselectivityusingcationicconjugatedpolymersandPicoGreen.Langmuir,25,43.
AAT Bioquest AAT Bioquest是一家位于美国的生物公司,前身为ABD Bioquest,总部位于加利福尼亚州。专门从事光学检测技术十多年,一直致力于光谱学检测领域技术的创新和突破。其独特的光学检测技术,综合了化学、生物学和信息学等各个领域的研究,引领了比色、荧光和发光技术新一代光学探针的浪潮。AAT Bioquest在全球拥有强大的经验丰富的专业分销商网络,为从小型研究机构到《财富》500强企业的各类客户提供卓越的产品和定制服务。
美国AATBioquestInc.(前身是ABDBioquest,Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域,包括吸收(颜色),荧光和发光技术。AATBioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学,免疫学,细胞生物学和分子生物学等领域。AATBioquest会不断介绍新产品,快速的丰富各个领域的产品。
1)我们提供反应荧光探针和发光探针,生物素和端粒酶能够应用于标记药物小分子和生物聚合物,如蛋白、核酸以及其他碳水化合物;2)我们研究并生产荧光和发光探针用于检测蛋白,核酸和活细胞。3)我们不断的推出新型的荧光和发光探针用于检测多种酶,特别是检测水解酶和氧化还原酶类;4)我们致力于开发用于信号转导研究的试剂;5)我们提供生理和神经探针,特别是钙离子指示剂和膜电位探针。
作为AATBioquestInc.的中国区域代理,艾美捷科技为中国客户提供光谱学检测领域,包括吸收(颜色),荧光和发光技术等全系列解决方案。我们也将一如既往更加努力为国内用户提供快捷、方便的高质量产品,同时更为您售前售后全面技术支持。
AATBioquest,Inc.(formerlyABDBioquest,Inc.)develops,manufacturesandmarketsbioanalyticalresearchreagentsandkitstoscientistsengagedinlifesciencesresearch,diagnosticR&Danddrugdiscovery.Wespecializeintheareaofphotometricdetectionsincludingabsorption(color),fluorescenceandluminescencetechnologies.TheCompany"sproductsenablescientistsandbiomedicalresearcherstobetterunderstandbiochemistry,immunology,cellBIOLOGyandmolecularbiology.AATBioquestconstantlyintroducesnewproducts,andoffersarapidlyexpandinglistofproductsthataregroupedintoseveralproductlines.
1)Ourreactivefluorescentandluminescentprobes,biotinsandtagenzymesareusedforlabelingsmalldrugmoleculesandbiopolymers,e.g,proteins,nucleicacidsandcarbohydrates;2)Wedevelopfluorescentandluminescentprobesfordetectingproteins,nucleicacidsandlivecells;3)Weconstantlyintroducenovelfluorescentandluminescentprobesfordetectingvariousenzymes,inparticular,hydrolyticandredoxenzymes;4)Wefocusondevelopingreagentsforsignaltransductionresearch;and5)Wealsoofferphysiologicalandneurologicalprobes,e.g.,calciumindicatorsandmembranepotentialprobes.
Besidesthestandardcatalogproductswealsooffercustomservicetomeetyourspecialresearchneeds.Ourcurrentservicesincludecustomsynthesisofcolorimetric,fluorescentandluminescentprobes,customdevelopmentofbiochemical,cell-basedanddiagnosticassaysandcustomscreeningofyourcompoundlibrariesagainstyourdefinedtargetsusingourvalidatedHTSassays.
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
谢谢谢
只要引物设计的可以在基因组上做,那么理论上是没问题的!
只需要用RT-PCR荧光定量试剂盒里面的第二个,专门用来做Real-timePCR的那些试剂就可以了!
如果要是直接注射,就不知道了
试剂盒里有详细的说明书,告诉你样品需要多少量,每个试剂需要加入多少量,和详细的实验步骤,一般买来就可以用,不用人教。
所以你问一个样需要多少量是没法回答的,测定过程是要加很多种试剂的。
定性ELISA只能粗略表示样本待测物含量在某个特定值以上或以下,定性ELISA通常设置阳性对照(P)、和阴性对照(N),结果分别用“阴性”和“阳性”表示。ELISA定性测定的“阴性”和“阳性”的判断标准是试剂盒所确定的阳性判断值,即cut-off值。
定量ELISA是通过一系列不同已知浓度的标准品所对应的OD值做出标准曲线,然后将样本的OD值代入标准曲线,计算出样本中待测物的含量。百特纯大分子Meretciel的定量ELISA试剂盒还可以。
暂无品牌问答