EvaGreen®dyeisagreenfluorescentnucleicaciddyewithfeaturesthatmakethedyeusefulforseveralapplicationsincludingqPCRandDNAmeltcurveanalysis,real-timemonitoringofthermophilichelicase-dependentamplification(tHDA),andcapillarygelelectrophoresis.Thedyeisessentiallynonfluorescentbyitself,butbecomeshighlyfluorescentuponbindingtodsDNA.TheDNA-bounddyehasexcitationandemissionspectraveryclosetothoseoffluorescein(FAM)orSYBR®GreenI,makingitreADIlycompatIBLewithinstrumentsequippedwiththe488nmargonlaseroranyvisiblelightexcitationwithwavelengthintheregion.EvaGreen®dyeisextremelystableboththermallyandhydrolytically,providingconvenienceduringroutinehandling.EvaGreen®dyeisnonmutagenicandnoncytotoxicbecauseitisimpermeabletocellmembranes,unlikeSYBR®GreenI,whichenterscellrapidlyandisknowntobeapowerfulmutation-enhancer(Ohta,etel.Mutat.Res.492,91(2001)).
TheuniquepropertiesofEvaGreen®dyehavemadeitparticularlyusefulinquantitativereal-timePCR(qPCR)applications.ComparedwiththewidelyusedSYBR®GreenI,EvaGreen®dyeisgenerallylessinhibitorytowardPCRandlesslikelytocausenonspecificamplification.Asaresult,EvaGreen®dyecanbeusedatamuchhigherdyeconcentrationthanSYBR®GreenI,resultinginahighresolutionsignalfordropletdigitalPCR,real-timePCRandmeltcurveanalysis.
Alsoseeourready-to-useFastEvaGreen®qPCRMasterMixandFastPlusEvaGreen®qPCRMasterMixes.
EvaGreendyeandapplicationsarecoveredunderUSpatentnos.7,803,943and7,776,567andpendinginternationalpatents.
Features:
LowPCRinhibition
ExhibitsmuchlessPCRinhibitionthanSYBR®GreenIviaasmart“release-on-demand”DNA-bindingtechnology
HighlySensitive
LowPCRinhibitionofthedyepermitsahigherdyeconcentrationtobeusedformuchgreaterqPCRandmeltcurveanalysissignals
Nonmutagenicandnoncytotoxic
NonmutagenicandnoncytotoxicbystandardAmestest;completelyimpermeabletocellmembranes
CompatiblewithfastPCRprotocols
MinimalinterferencetoPCRmakesitpossibletosignificantlyshortenthechainextensiontime
CompatiblewithmultiplexPCR
Nodyemigrationfromamplicontoampliconwhenusedattherecommendedconcentration
UnsurpassedthermalstABIlity,chemicalstabilityandphotostability
NodetectabledyedecompositioninPCRbufferat95-100°Cfor48hours;highlystableundereitheralkalineoracidiccondition;withstandsrepeatedfreeze-thawcycles
SpectrallysimilartoSYBR®GreenI
CompatiblewithallmajorbrandqPCRinstrumentsandenzymesystems
References
Mao,etal.CharacterizationofEvaGreenDyeandtheimplicationofitsphysicochemicalpropertiesforqPCRapplications.BMCBiotechnology7,76(2007).
EvaGreenindropletdigitalPCR
1.LabChip(2010),doi:10.1039/c004521g
2.TheRoyalSocietyofChemistry(2014),doi:10.1039/c3an02334f
3.AnalyticalChemistry(2014),doi:10.1021/ac403843j
4.AnalyticalChemistry(2013),dx.doi.org/10.1021/ac4030413
5.TheJournalofAmericanSociety(2011),dx.doi.org/10.1021/ja2060116
6.AnalyticalChemistry(2010),doi:10.1021/ac902510u
7.AnalyticalChemistry(2013),doi:10.1021/ac403061n
8.NucleicAcidsResearch(2013),doi:10.1093/nar/gkt684
DownloadalistofSelectedEvaGreenReferences.
