Pleaseseethe EpiQuik™MethylatedDNAImmunoprecipitationKit ifyouareworkingwithnuclearextractasthestartingmaterial.

The Methylamp™MethylatedDNACapture(MeDIP)KitisacompletesetofoptimizedreagentstoenrichandcapturemethylatedDNAfragmentsinaconvenientmicroplate-basedformat.Themethod,methylatedDNAimmunoprecipitation(meDIP),usesa monoclonalantibodyspecificto5-methylcytosine toimmunoprecipitatemethylatedgenomicDNA.Theenrichedmethylatedfractionscanthenbeusedforgene-specificDNAmethylationanalysisonagenomewidescale.The Methylamp™MethylatedDNACaptureKitissuitableforcombiningthespecificityofenrichedmethylatedDNAwithqualitativeandquantitativePCR,trADItionalsequencingandnext-generationsequencing,DNAmicroarrays,aswellasSouthernblot.Thekithasthefollowingadvantagesandfeatures:

• HighlyefficientenrichmentofmethylatedDNA:>98%.
• Extremelyfastprocedure,whichcanbefinishedwithin3hours.
• Strip-wellmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
• ColumnsforDNApurificationareincluded,savingtimeandreducinglabor.
• CompatiblewithallDNAamplification-basedapproaches.
• Simple,reliable,andconsistentassayconditions.

BackgroundInformation
MeDIPisagenome-wide,largescaletechniquetoenrichmethylatedDNAbyisolatingmethylatedDNAfragmentswitha5-methylcytosineantibody.Thistechniquehasplayedanimportantroleinhelpingtoidentifyabnormallymethylatedgenesindiseasessuchascancer.WithMeDIP,theimmunoprecipitatedfractionsofmethylatedDNAcanbedeterminedbyPCRtoevaluatethemethylationstateofindividualregions.Additionally,thepurifiedfractionscanbeinputtohighthroughputDNAdetectionmethodssuchasDNAmicroarrays(MeDIP-chip)andnext-generationsequencing(MeDIP-seq),becomingausefulapproachformethylome-levelanalysisandfordevelopingcomprehensiveprofilesofDNAmethylationonagenome-widescale.

Principle&Procedure
ThemostcommonandcrucialexperimentalfactorofMeDIPthatmayaffectitsaccuracyinvolvesthequalityandcross-reactivityofthe5-methylcytosineantibodyusedintheprocedure.EpigentekovercamesuchalimitationthroughextensiveresearchanddevelopmenttocreateahighlyefficientMeDIPkitbyapplyinganon-cross-reactive5mCantibody.Inthekit’sassay,DNAisshearedandthenaddedintoamicroplatewellormultiplewellsinahighthroughputformat.AhighqualityChIP/MeDIP-gradeantibodyspecifictomethylcytosineisthenusedtocapturemethylatedDNAinthewells.Theantibody-capturedmethylatedDNAisreleasedandisfinallypurifiedthroughtheincludedspincolumns.TheelutedmethylatedDNAcannowbeusedinvariousdown-streamapplications.

MC1(AntibodyBuffer)
MC2(ReactionBuffer)
MC3(WashBuffer)
MC4(DNAReleaseBuffer)
MC5(BindingBuffer)
MC6(ElutionBuffer)
NormalMouseIgG(1mg/ml)*
Anti-5-Methylcytosine(1mg/ml)*
ProteinaseK(10mg/ml)*
8-WellAssayStrips(withFrame)
8-WellStripCaps
F-SpinColumn
F-CollectionTube
UserGuide

*Formaximumrecoveryoftheproducts,centrifugetheoriginalvialafterthawingpriortoopeningthecap.