| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 501/520 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | NucleicAcidDetection DNADetection |
| Related | LabelingCells FluorescenceImaging BiochemicalAssays |
1.Preparing1XAssayBuffer
Preparea1XAssaybufferbydilutingtheconcentratedbuffer20-foldwithsterile,distilled,DNase-freewater.
2.PreparingHelixyteGreen™workingsolution
PrepareHelixyteGreen™workingsolutionbymakinga200-folddilutionoftheconcentratedDMSOsolutionin1Xassaybuffer.Forexample,toprepareenoughworkingsolutiontoassay10samplesina2mLfinalvolume,add50µLofHelixyteGreen™(ComponentA)into10mLofAssayBuffer(fromStep1). Protecttheworkingsolutionfromlightbycoveringitwithfoilorplacingitinthedark.
Note1:Werecommendpreparingthissolutioninaplasticcontainerratherthanglass,asthedyemayadsorbtoglasssurfaces.
Note2:Forbestresults,thissolutionshouldbeusedwithinafewhoursofitspreparation.
3.PrepareserialdilutionsofdsDNAstandard(0to2µg/mL):
3.1 Forhighrangestandardcurve:add30μLof100μg/mLdsDNAstocksolution(ComponentC)to1.47mLof1XAssaybuffer(FromStep1)tohave2000ng/mLdsDNAsolution,andthenperform1:2and1:10serialdilutionstoget1000,100,10,1and0ng/mL.
3.2 Forlowrangestandardcurve:add40μLof2μg/mLdsDNAstocksolution(FromStep3.1)to1.56mLof1XAssaybuffer(FromStep1)tohave50ng/mLdsDNAsolution,andthenperform1:2and1:10serialdilutionstoget25,2.5,0.25,0.025and0ng/mL
4.RundsDNAassay:
4.1 Add1mLofHelixyteGreen™workingsolution(fromStep2)toeachcuvettecontaining1mLofthedsDNAstandard,blankcontrol,andtestsamplestomakethetotaldsDNAassayvolumeof2mL/cuvette.
4.2 Incubatethereactionatroomtemperaturefor5to10minutes,protectedfromlight.
4.3 MonitorthefluorescenceincreasewithaspectroflurometeratEx/Em=490/525nm.
Note:Tominimizephotobleachingeffects,keepthetimeforfluorescencemeasurementconstantforallsamples.
4.4 Thefluorescenceinblankwells(withtheAssaybufferonly)isusedasacontrol,andissubtractedfromthevaluesforthosecuvetteswiththedsDNAreactions.TheDNAconcentrationofthesamplesisdeterminedfromthestandardcurvegeneratedinDNAStandardCurve.
| 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。
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1.模板提取（一般为RNA）：Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定：分光光度计或NanoDrop
3.逆转录：逆转录试剂盒（或者一步法试剂盒），这一步可以用普通PCR做，也可以用水域做。
4.荧光定量PCR试剂：通常有用SYBR Green Mix做的，但是这里建议你用EvaGreen做，灵敏度和平行性都要好于SYBR Green，并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他：除了以上的那些还需要离心管、PCR管或板（Axygen反应比较好）、移液枪等，暂时就想到这么多。
1.模板提取（一般为RNA）：Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定：分光光度计或NanoDrop
3.逆转录：逆转录试剂盒（或者一步法试剂盒），这一步可以用普通PCR做，也可以用水域做。
4.荧光定量PCR试剂：通常有用SYBR Green Mix做的，但是这里建议你用EvaGreen做，灵敏度和平行性都要好于SYBR Green，并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他：除了以上的那些还需要离心管、PCR管或板（Axygen反应比较好）、移液枪等，暂时就想到这么多。
MicroBCA试剂测定范围是0.5-20μg/ml
楼主自己看看经销商价格
临床实验室对商品定量试剂盒分析性能的验证.pdf(17431.6k)
如果要是直接注射，就不知道了
