TheMethylamp™One-StepDNAModificationKitisacompletesetofessentialcomponentswhichenablestheexperimentertoperformDNAmethylationanalysisusingEpigentek"suniquelysimplifiedandstreamlinedbisulfitemethod.Theentireprocedurecanbecompletedwithinamere1hourand45minutesandproducesfarsuperiorresultsthananycompetitorkits.TheMethylamp™One-StepDNAModificationKitissuitableforMS-PCR,realtimeMS-PCR,methylationsequencing,andpyrosequencing,aswellasmethylationmicroarray.Thekithasthefollowingadvantages:

• Thenumberonefastestprocedureavailable.Theentireexperimentcanbefinishedwithin1hourand45minutes.
• Completelyconvertsunmethylatedcytosineintouracil:modifiedDNA>99.9%.
• ThelowestdegradationofDNAinthemodificationprocess:morethan90%ofDNAlosscanbeprevented.
• ThelowestrequirementofstartingDNAformodification:only50pgor20cells.
• Extremelysimple,reliable,andconsistentmodificationconditions.

Principle&Procedure
TheMethylamp™One-StepDNAModificationKitcontainsallreagentsrequiredforbisulfiteconversiononaDNAsample.DNAisdenaturedbyheating,whichallowsDNAdenaturationandbisulfitemodificationtobecarriedoutsimultaneously.Inthemodificationprocess,bisulfitereagentreactsspecificallywithsingle-strandedDNA,therebydeaminatingcytosineandcreatingauracilresidue.TheuniqueDNAprotectionreagentscontainedinthemodificationbuffercanpreventthechemicalandThermophilicdegradationofDNAinthebisulfitetreatment.Thenon-toxicmodifiedDNAcapturebufferenablesDNAtotightlybindtothecolumnfilter,thusDNAcleaningcanbecarriedoutonthecolumntoeffectivelyremoveresidualsodiumbisulfiteandsalts.ModifiedDNAthencanbeelutedandstablystoredat-20°Cforupto2months.

Fig.2. ComparativeoverviewforusingtheMethylampOne-StepDNAModificationKit.

 
Fig.1.SchematicprocedureforusingtheMethylampOne-StepDNAModificationKit.