| InputType: | DNA |
| ResearchArea: | DNAMethylation |
| TargetApplication: | SampleModification |
| VesselFormat: | Columns/Tubes |
| 100%Guarantee: | 6months |
The Methylamp™WholeCellBisulfiteModificationKit isaninnovativeanduniquesetofessentialcomponentswhichenablestheexperimentertoperformDNAmethylationanalysisandmodifyDNAdirectlyfromcellsortissuesusingEpigentek"suniquelysimplifiedandstreamlinedbisulfitemethod.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
