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NEB/Monarch® PCR & DNA Cleanup Kit (5 μg)/T1030S/250 preps
  • NEB/Monarch® PCR & DNA Cleanup Kit (5 μg)/T1030S/250 preps

NEB/Monarch® PCR & DNA Cleanup Kit (5 μg)/T1030S/250 preps

价格: ¥5460.00 市场价: 9100.00
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货号: T1030S-250preps
品牌: NEB
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    • Description:

       
      TheMonarchPCR&DNACleanupKitrapidlyandreliablypurifiesupto5μgofconcentrated,high-qualityDNAfromPCRandotherenzymaticreactions.Thekitutilizesabind/wash/eluteworkflowwithminimalincubationandspintimes.Thecolumnsensurezerobufferretentionandnocarryoverofcontaminants,enablingelutionofsampleinvolumesaslowas6μl.Thebuffersprovidedhavebeenoptimized,anddonotrequiremonitoringofpH.ElutedDNAisreadyforuseinrestrictiondigests,DNAsequencing,ligationandotherenzymaticmanipulations.DesignedwithsustainABIlityinmind,MonarchkitsusesignificantlylessplasticandresponsIBLy-sourced,recyclablepackaging.TheprotocolcanalsobemodifiedtoenablethepurificationofsmallerDNAfragments,includingoligonucleotides.

      Viewourvideosonprotocols,tips,andrecyclingMonarch.

       
      APPLICATIONS
      PCRcleanupDNAfromPCRreactionscanbepurifiedafteramplificationtoremovepolymerases,primers,detergents,dNTPs,etc.
      EnzymaticreactioncleanupModifyingenzymessuchasligases,kinases,nucleases,phosphatasesareefficientlyremovedallowingefficientdesaltingandconcentrationoftheDNAsample.
      CDNAcleanupDNA/RNAcomplexescanbepurifiedpost-reversetranscription/amplificationtoenableremovaloftheRTandpolymeraseaswellasnucleotides.
      LabelingcleanupUnincorporatedrADIolabeledorfluorescentlylabelednucleotidescanberemovedfromtheDNAsubstrate
      PlasmidcleanupPlasmidprepsfromunknownsourcesmaycontaininhibitorsandunwantedcontaminants.Purificationandconcentrationcanbeeasilyachievedusingthiskit.
      OligonucleotidecleanupssDNAoligonucleotides(≥18nt)anddsDNAfragments(≥ 15bp)canbepurifiedusingtheOligonucleotideCleanupProtocol.


      MonarchDNACleanupColumnDesign
      Monarch DNA Cleanup Column Design

      Monarchcolumnsaredesignedforperformance
      Monarch columns are designed for performance. Monarch columns are designed without a frit, which eliminates buffer retention and the risk of carryover contamination, providing fast, worry-free DNA purification.
      Monarchcolumnsaredesignedwithoutafrit,whicheliminatesbufferretentionandtheriskofcarryovercontamination,providingfast,worry-freeDNApurification.

       Advantages:
      • Eluteinaslittleas6μl
      • Preventbufferretentionandsaltcarry-overwithoptimizedcolumndesign
      • Savetimewithfast,user-friendlyprotocols
      • NoneedtomonitorpH
      • Buffersandcolumnsavailableseparately
      • Significantlylessplasticusedwhencomparedwithotherkits 
      • Responsibly-sourcedandrecyclablepackaging 

      Specifications

      DNASampleType:DNAfromPCRandotherenzymaticreactions(e.g.,restrictiondigests,kinasereactions,ligations).*
      BindingCapacity:upto5μg
      DNASizeRange:~50bpto25kb**
      TypicalRecovery:DNA(50bpto10kb):70–90%/ DNA(11–23kb):50–70%
      ElutionVolume:≥6μl
      Purity:A260/280>1.8andA260/230>1.8
      ProtocolTime:5minutesofspinandincubationtime
      CompatibleDownstream
      Applications:      		ligation,restrictiondigestion,labelingandotherenzymatic
      manipulations,libraryconstructionandDNAsequencing.

      *ssDNAordsDNAoligonucleotidesfromenzymaticreactionscanalsobepurifiedusingtheOligonucleotideCleanupProtocol.
      **DNA≥15bpto25kb(dsDNA)andDNA≥18ntto10kb(ssDNA)canalsobepurifiedusingtheOligonucleotideCleanupProtocol.
       

      MonarchPCR&DNACleanupKit(5µg)Protocol 
      Monarch PCR & DNA Cleanup Kit (5 µg) Protocol

      MonarchPCR&DNACleanupKit(5μg)performsequivalentlytotheleadingsupplier
      Monarch PCR & DNA Cleanup Kit (5 μg) performs equivalently to the leading supplier. Preps were performed according to recommended protocols. 1 μg of a 3 kb DNA fragment was incubated with 1 μM primers and OneTaq® Quick-Load® 2X Master Mix (NEB #M0486). DNA was eluted in 20 μl (NEB) and 40 μl (Qiagen) Elution Buffer. Half of the total elution volume was digested with 5 units of DraIII-HF® (NEB #R3510). The digest and the unused portion of the elution were resolved on a 1% w/v agarose gel along with a representative sample of the starting material.
      Prepswereperformedaccordingtorecommendedprotocols.1μgofa3kbDNAfragmentwasincubatedwith1μMprimersandOneTaq®Quick-Load®2XMasterMix(NEB#M0486).DNAwaselutedin20μl(NEB)and40μl(Qiagen)ElutionBuffer.Halfofthetotalelutionvolumewasdigestedwith5unitsofDraIII-HF®(NEB#R3510).Thedigestandtheunusedportionoftheelutionwereresolvedona1%w/vagarosegelalongwitharepresentativesampleofthestartingmaterial. 
      MonarchPCR&DNACleanupKit(5μg)removeslowmolecularweightprimersfromdsDNAsamples
      Monarch PCR & DNA Cleanup Kit (5 μg) removes low molecular weight primers from dsDNA samples. Three independent amplicons (267 bp, 520 bp, 1003 bp) were spiked with two oligonucleotides (16-mer, 24-mer) to a final concentration of 1 μM. Half of each mix was purified with the Monarch PCR & DNA Cleanup Kit (5 μg) following the included protocol. Equivalent fractions of the original mixture and the eluted material were resolved on a 20% TBE acrylamide gel at 100V for one hour and stained with SYBR Green II.
      Threeindependentamplicons(267bp,520bp,1003bp)werespikedwithtwooligonucleotides(16-mer,24-mer)toafinalconcentrationof1μM.HalfofeachmixwaspurifiedwiththeMonarchPCR&DNACleanupKit(5μg)followingtheincludedprotocol.Equivalentfractionsoftheoriginalmixtureandtheelutedmaterialwereresolvedona20%TBEacrylamidegelat100VforonehourandstainedwithSYBRGreenII.

      LearnMoreAboutMonarch
      • NEBMonarch.com
      • Optimizeyourresultswithouruniquecolumndesign
      • EnhanceyourDNApurificationexperience
      • FeelgoodaboutchoosingMonarch
      • ChooseMonarchkitsforpurevalue

      KitComponents

      Thefollowingreagentsaresuppliedwiththisproduct:

      Storeat(°C)Concentration
      Monarch®DNACleanupBindingBuffer251X
      Monarch®DNAWashBuffer255X
      Monarch®DNAElutionBuffer251X
      Monarch®DNACleanupColumns(5μg)25

      Notes:

      Thekitshouldbestoredatroomtemperature.Alwayskeepbufferbottlestightlyclosedandkeepcolumnssealedintheenclosedzip-lockbag.Forinformationregardingthecompositionofbuffers,pleaseconsulttheSafetyDataSheets.Properlaboratorysafetypracticesshouldbeemployed,includingtheuseoflabcoats,glovesandeyeprotection.
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