PMAisahighaffinityphotoreactiveDNAbindingdyedevelopedbyBiotiumforviABIlityPCR(40019).PMAxx™isournewandimprovedviabilityPCRdye(40069).Thedyesareweaklyfluorescentbythemselvesbutbecomehighlyfluorescentuponbindingtonucleicacids.Uponphotolysis,thedyeformsastablecovalentbondwiththeDNA,thusresultinginpermanentDNAmodification.Thedyeiscellmembrane-impermeableandthuscanbeusedtoselectivelymodifyDNAfromdeadcellswithcompromisedmembraneintegrity,whileleavingDNAfromviablecellsintact.PMAandPMAxx™ inhibitPCRamplificationofmodifiedDNAtemplates.Consequentlythedyeisusefulintheselectivedetectionofviablepathogeniccellsbyquantitativereal-timePCR.
PMAEnhancerforGramNegativeBacteriawasdesignedtoimprovePMA-andPMAxx™-mediateddiscriminationbetweenliveanddeadgram-negativebacteria.PMAEnhancerisprovidedasa5Xsolution,andisaddedtoasamplebeforetheadditionofdye.Whenasequencefromagram-negativebacteriaisamplifiedbyPCR,samplespre-treatedwithEnhancershowadecreaseinthesignalfromdeadcells,withnochangeinthesignalfromlivecells.Thus,PMAorPMAxx™plusEnhanceristheoptimalwaytoperformviabilityPCRongram-negativebacteria.
Note:PMAEnhancerisnotintendedforusewhengram-positivebacteriaaretobedetected.PMAEnhancermayadverselyaffecttheamplificationoflivecellDNAfromgram-positivebacteria.
ForphotoactivationofPMA,PMAxx™orEMAdye,werecommendtheuseofBiotium’sPMA-Lite™LEDPhotolysisDevice(E90002),whichisdesignedtoconductphotolysisundercontrolledconditions.
AlsoseeourPMAReal-TimeBacterialViabilityKits.
MaterialsfromBiotiumaresoldforresearchuseonly.
References
1.Nocker,A.,etal.Comparisonofpropidiummonoazidewithethidiummonoazidefordifferentiationoflivevs.deadbacteriabyselectiveremovalofDNAfromdeadcells.J.Microbiol.Meth.67(2),310-320(2006).
2.Nocker,A.,etal.MolecularmonitoringofdisinfectionefficacyusingpropidiummonoazideincombinationwithquantitativePCR.J.Microbiol.Meth.70(2),252-260(2007).
3.Fittipaldi,M.,etal.ProgressinunderstandingpreferentialdetectionoflivecellsusingviabilitydyesincombinationwithDNAamplification.J.Microbiol.Meth.91(2),276-289(2012).
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