ObtainhighyieldsofqualityDNAfromyeastwiththiseasytousekit
- Fast:ObtainpurifiedDNAinlessthan40minutes
- FlexIBLe:PurifyDNAfromawidevarietyofyeastspecies,includingCandida,Saccharomyces,Pichia,andSchizosaccharomyces,andfilamentousfungisuchasAspergillus2andPenicillium
- HighQualityDNA:PurifyhighmolecularweightgenomicDNAthatisreadytouseinmanymolecularBIOLOGyapplications,includingPCRamplification,restrictionendonucleasedigestion,Southernblotting,genomiclibrarypreparationandfungalidentificationandtyping
- HighYields:PurifymoreyeastgenomicDNAthanothercommerciallyavailablekits
- Simple:Noenzymaticlysis,beadbeating,columns,phenolorotherorganicsolventsareused
TheMasterPure™YeastDNAPurificationKitenablesefficient,high-yieldpurificationofhigh-molecular-weightDNAfromyeastandotherfungi.Theprotocolinvolvesnonenzymaticcelllysisat65°C,followedbyremovalofproteinbyprecipitation,andnucleicacidprecipitationandresUSPension.Nolyticase,proteolyticenzymes,orbead-beatingareusedintheprocedure.YeastgenomicDNAyieldsusingtheMasterPureKitaremuchhigherthanyieldsobtainedwithothercommerciallyavailablekits(Fig.1).1Theprotocolcanbeeasilyadjustedforlargerorsmallersamples,includingsingleyeastcolonies.1Therecoverednucleicacidcanbeuseddirectlyinmostapplications,includingPCRamplification.
References
- Hoffman,L.andMoan,E.(1998)EpicentreForum5(4),1.
- Jin,J.,Lee,Y.-K.andWickes,B.L.(2004)J.ClinMicrobiol.42,4293.
Figure1.ComparativeyieldsofDNAfromtheMasterPure™YeastDNAPurificationKitandcompetitorkits.DNAwasquantitatedspecificallywithHoechstfluorescentdye33258,whichgivesminimalfluorescencewithRNA.ThedatarepresenttheaverageDNAyieldsdeterminedbyfluorometryfromtwoexperimentswithS.cerevisiaeandC.albicans.TheMasterPureKitproducedupto17timesmoreDNAfromC.albicansand12timesmoreDNAfromS.cerevisiaethanotherkits. |
Figure2.SizedistributionofDNAproducedfromtheMasterPure™Kit.Foreachkitindicated,a500-ngaliquotofpurifiedyeastDNAwasanalyzedbypulse-fieldgelelectrophoresisona1%agarosegel.Thegelwasstainedwithethidiumbromide.ThemajorityofDNAisolatedwiththeMasterPureKitwasestimatedtobeinthe40-to50-kbsizerange,whiledegradationtosmallerfragmentswasobservedwithotherkits.LaneM,lamBDaDNAladder;Lane4,T7DNA(40kb). |