| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 500/525 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | NucleicAcidDetection RNADetection |
| Related | CellFunctionalAnalysis FluorescenceImaging BiochemicalAssays |
1.Preparing1Xassaybuffer
Preparea1Xassaybufferbydilutingtheconcentratedbuffer20X(ComponentB)withsterile,distilled,nuclease-freewater.
2.PreparingStrandBrite™Greenworkingsolution
PrepareStrandBrite™Greenworkingsolutionbymakinga200-folddilutionoftheconcentratedDMSOsolutionin1Xassaybuffer.Forexample,toprepareenoughworkingsolutiontoassay10samplesina2mLfinalvolume,add50µLofStrandBrite™Green(ComponentA)into10mLof1Xassaybuffer(fromStep1). Protecttheworkingsolutionfromlightbycoveringitwithfoilorplacingitinthedark.
Note1:Werecommendpreparingthissolutioninaplasticcontainerratherthanglass,asthedyemayadsorbtoglasssurfaces.
Note2:Forbestresults,thissolutionshouldbeusedwithinafewhoursofitspreparation.
3.PrepareserialdilutionsofRNAstandard(0to1µg/mL):
Add20μLof100μg/mLRNAstocksolution(ComponentC)to1.98mLof1Xassaybuffer(FromStep1)tohave1µg/mLRNAsolution,andthenperform1:3serialdilutionstogetapproximately1000,300,100,30,10,3,1,and0g/mL.
Note:UnusedRibosomalRNAStandard(ComponentC)shouldbedividedintosingleusealiquotsinnuclease-freeplasticvialsandstoredat-20ºC.
4.RunRNAassay:
4.1 Add1mLofStrandBrite™Greenworkingsolution(fromStep2)toeachcuvettecontaining1mLoftheRNAstandard,blankcontrol,andtestsamplestomakethetotalRNAassayvolumeof2mL/cuvette.
4.2 Incubatethereactionatroomtemperaturefor2to5minutes,protectedfromlight.
4.3 MonitorthefluorescenceincreasewithaspectrofluorometeratEx/Em=490/545nm.
Note:Tominimizephotobleachingeffects,keepthetimeforfluorescencemeasurementconstantforallsamples.
4.4 Thefluorescenceinblankwells(withtheassaybufferonly)isusedasacontrol,andissubtractedfromthevaluesforthosecuvetteswithRNAstandardortestsamples.TheRNAconcentrationofthesamplearedeterminedaccordingtotheRNAstandardcurve.
| References&Citations |  PrinterFriendlyVersion |
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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.
