快速分离和纯化小RNA分子(<200 nt)方案_miRNA
货号: | 325-106;325-150 |
英文名: | Hybrid-RmiRNA |
保存条件: | Roomtemperature(15~25℃)/4℃(RiboEx) |
Cat.No. | ProductDescription | Size |
325-106 | Hybrid-RTM miRNA | 6 |
325-150 | Hybrid-RTM miRNA | 50 |
Description
Inrecentyears,interestinsmallRNA,suchassiRNAandmiRNAwhicharerelatedtoresearchofgeneregulation,hasexpanded.TherearemanycommercialkitsfortotalRNApreparation,butmostofthesearefocusedonpreparationoflargeRNAlongerthan200nucleotides.BecausebothsiRNAandmiRNAarebetween15~30nucleotidesinlength,theneedofspeciallyoptimizedkitforsmallRNA(<200nucleotides)isgrowingrapidly.Hybrid-RTM miRNAisdesignedforpurificationoflargeandsmallRNAseparatelyfromculturecellsoranimaltissuesandco-purificationinasingletubeisalsoavailablebymodifiedprotocol.ThiskitutilizesthelysismethodofRiboExTM whichhasapowerfulabilityoflysisandthepurificationmethodbasedonglassfibermembranetechnology.SamplesarehomogenizedinRiboExTM,amonophasicsolutioncontainingphenolandguanidiumsalt,whichrapidlylysecellsandinactivatesnucleases.Additionofchloroformbringsaboutaseparationofthelysateintoaqueousandorganicphases.TotalRNAlocatesintheaqueousphasewhileDNAandproteinremainintheinterphaseandorganicphase.LargeandsmallRNAintheaqueousphaseisselectivelyboundtocolumntypeBandtypeWrespectively.ThecolumntypeBselectivelyadsorbstheRNAlargerthan200nucleotidesinlength,whilethecolumntypeWspecificallyholdstheRNAsmallerthan200nucleotidesinlength.TopurifylargeRNA,theaqueousphaseismixedwithethanolandthemixtureisappliedtoacolumntypeB.Aftercentrifugation,largeRNAisboundtomembraneandthemixturecontainingsmallRNAgoesintocollectiontubethroughthemembrane.Themembraneiswashedawaybytwowashbuffer(SW1andRNW)andpurifiedlargeRNAiselutedfromthemembranebyRNase-freewater.TopurifysmallRNA,thepass-throughcomefromthebindingoflargeRNAismixedwithethanolandthenappliedtoacolumntypeW.AfterwashingwithbufferRBWandRNW,smallRNAiselutedbyRNase-freewater.TheprocedureofHybrid-RTM miRNAtakesonly30minutesforcompletepreparationsofpureRNA.ThepurifiedRNAissuitablefortheisolationofPolyA+RNA,northernblotting,dotblotting,invitrotranslation,cloning,RT-PCR,RPAandotheranalyticalprocedures.
FeaturesandBenefits
■Preparationtime:~30minutes
■Stableandconsistentyield
■Highpurityandyield
■PerfectseparationofsmallRNAfragment
■Samplesize:Upto50mgtissueorupto1x107 culturedcells
■Recoveryrange:LargeRNA:>200nucleotides
SmallRNA:<200nucleotides
■Instantuse:Noneedofadditionalmaterials
■Noethanolprecipitation
■NoGenomicDNAcontamination
■Readyforuseinnorthernblotting,dotblotting,invitrotranslation,cloning,RT-PCR,RPAandother analyticalprocedures
Procedure
温馨提示:不可用于临床治疗。
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发布于 : 2018-03-04
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