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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *High Sensitivity*/17651/200 Tests
产品编号:17651
市  场 价:¥56560.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$2828.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *High Sensitivity*/17651/200 Tests
商品介绍
Overview
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Ex/Em(nm)501/520
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryNucleicAcidDetection
DNADetection
RelatedLabelingCells
FluorescenceImaging
BiochemicalAssays
Helixyte™GreendsDNAQuantitationAssayKitcanbeusedforselectivelydetectingaslittleas25pg/mlofdsDNAinthepresenceofssDNA,RNA,andfreenucleotides.Helixyte™GreenexhibitslargefluorescenceenhancementuponbindingtodsDNA.Theassayislinearoverthreeordersofmagnitudeandhaslittlesequencedependence,allowingyoutoaccuratelymeasureDNAfrommanysources,includinggenomicDNA,viralDNA,miniprepDNA,orPCRamplificationproducts.Helixyte™GreendsDNAQuantitationAssayKitisafewmagnitudesmoresensitivethanUVabsorbancereADIngs.ItisspecificfordsDNAinthepresenceofequimolaramountsofRNA.Thekitisrobustwithamixandreadformat.Itcanbeusedwithabenchtopfluorometerorahand-heldfluorescencemeter(e.g.,qubitfluorometer).ThiskitisanexcellentreplacementforQuant-iT™PicoGreen®dsDNAAssayKit(Quant-iT™andPicoGreen®arethetrademarksofInvitrogen).

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

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
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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.


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