Epigentek/Methylamp Whole Cell Bisulfite Modification Kit/P101680/80 samples

The Methylamp™WholeCellBisulfiteModificationKit isaninnovativeanduniquesetofessentialcomponentswhichenablestheexperimentertoperformDNAmethylationanalysisandmodifyDNAdirectlyfromcellsortissuesusingEpigenteksuniquelysimplifiedandstreamlinedbisulfitemethod.Theentireprocedurecanbecompletedwithinonly3hours.TheMethylamp™WholeCellBisulfiteModificationKitisspecificallydesignedforDNAmethylationresearchusingminuteamountsofstartingmaterialsincludingcellsculturedin96-well/384-wellplates,tissuesectionsamples,microdissectionsamples,tissuebiopsyandearlyembryoniccells/oocytes. 

ModifiedDNAiselutedbyusingtheMethylamp™WholeCellBisulfiteModificationKitandissuitableforrealtimeMS-PCR.ItisalsosuitableforalltechniquescurrentlyusedfortheanalysisofDNAmethylation;includingconventionalMS-PCR,bisulfitesequencing,pyrosequencing,andmethylationmicroarray.IfyouusetheMethylamp™WholeCellBisulfiteModificationKitforMSP,thenumbersofPCRcyclesshouldbegreaterthan45. Thekithasthefollowingadvantages:

• DirectlymodifyDNAincellsortissues.
• Rapidstreamlined3hourprocedure.
• Completelyconvertsunmethylatedcytosineintouracil:modifiedDNA>99.5%.
• ThelowestdegradationofDNAinthemodificationprocess:morethan90%ofDNAlosscanbepreventedwithauniqueDNAprotectionbuffer.

Principle&Procedure
TheMethylamp™WholeCellBisulfiteModificationKitcontainsallreagentsrequiredforbisulfiteconversiondirectlyonacellortissuesample.ThekitallowsDNAtobeisolatedfromcellsortissues,denaturedandbisulfitemodifiedsimultaneouslyinsametubewiththespecificreactionbufferunderthethermodynamiccondition.Inthemodificationprocess,bisulfitereagentreactsspecificallywithsingle-strandedDNA,therebydeaminatingcytosineandcreatingauracilresidue.TheuniqueDNAprotectionreagentscontainedinthemodificationbuffercanpreventthechemicalandthermophilicdegradationofDNAinthebisulfitetreatment.Thenon-toxicmodifiedDNAcapturebufferenablesDNAtotightlybindtothecolumnfilter,thusDNAcleaningcanbecarriedoutonthecolumntoeffectivelyremoveresidualsodiumbisulfiteandsalts.ModifiedDNAcanthenbeelutedandstablystoredat-20°Cforupto2months.

StartingMaterials
Theamountofstartingmaterialsforeachmodificationcanbe100-20000cells,or1µg-100µgoftissue,or0.2-2mm² tissuesectionsamples.Foroptimalmodification,theamountshouldbe500-5000cells,or5-20µgoftissue,or0.5-1mm² tissuesectionsamples,respectively.

W1(DigestionPowder)
W2(DigestionSolution)
W3(CellCollectionBuffer)
W4(DNAModificationPowder)
W5(DNAModificationBuffer)
W6(BalanceBuffer)
W7(DNABindingBuffer)
W8(ModifiedDNAElution)
F-SpinColumn
F-CollectionTube
UserGuide