當前位置： NEB >>> NEB/NEBuilder? HiFi DNA Assembly Master Mix/E2621S/250 reactions

NEB/NEBuilder? HiFi DNA Assembly Master Mix/E2621S/250 reactions

價格: ￥30240.00 市場價: 50400.00

打印產品信息

貨號: E2621S-250reactions

品牌: NEB

規格

默認

數量

庫存(0)

特別提示

代購產品：無質量問題不接受退換貨，下單前請仔細核對信息。
下單后請及時聯系客服核對商品價格，訂單生效后再付款。

立即購買加入購物車

收藏商品

資深產品顧問

咨詢顧問

全國免費服務熱線

4000-520-616

自營商城一站式服務
廠家直采剔除溢價
品質甄選正品保證
嚴控流程只做188精品
極速物流如約送貨

詳情
使用說明
常見問題

- Description:
  
  NEBuilderHiFiDNAAssemblyMasterMixwasdevelopedtoimprovetheefficiencyandaccuracyofDNAassembly.ThismethodallowsforseamlessassemblyofmultipleDNAfragments,regardlessoffragmentlengthorendcompatibility.Thismethodhasbeenusedtoassembleeithersingle-strandedoligonucleotidesordifferentsizesofDNAfragmentswithvariedoverlaps(15–80bp).IthasutilityforthesyntheticBIOLOGycommunity,aswellasthoseinterestedinone-stepcloningofmultiplefragmentsduetoitseaseofuse,flexibilityandsimplemaster-mixformat.Thereactionincludesdifferentenzymesthatworktogetherinthesamebuffer(seeFigure1):
  - Theexonucleasecreatessingle-stranded3′overhangsthatfacilitatetheannealingoffragmentsthatsharecomplementarityatoneend(theoverlapregion)
  - Thepolymerasefillsingapswithineachannealedfragment
  - TheDNAligasesealsnicksintheassembledDNA?
  Theendresultisadouble-strandedfullysealedDNAmoleculethatcanserveastemplateforPCR,RCAoravarietyofothermolecularbiologyapplications,includingdirecttransformationofEcoli.
  
  NEBuilderHiFiDNAAssemblykitsareavailableinvariousformats:withNEB5-alphacompetentcells(CloningKit,NEB#E5520),asabundlewithNEB10-betacompetentcells(BundleforLargeFragments,NEB#E2623)andwithoutcompetentcells(MasterMix,NEB#E2621).NEB5-alphacompetentcellscellsareexcellentforroutineassembliesof15kborless.NEBrecommendsNEB10-betaCompetentE.coli(HighEfficiency,NEB#C3019)orNEB10-betaElectrocompetentE.coli(NEB#C3020)forassemblieslargerthan15kb.Iftheassembledgenescontainrepetitivesequences,NEBStableCompetentE.coli(NEB#C3040)shouldbeused.?
  
  NEBuilderhasbeenusedinvariousapplications,including:
  - Site-directedmutagenesis
  - ConstructionofansgRNA-Cas9expressionvector|Animation
  - Assemblyoflinearyeastexpressioncassettes
  TohelpselectthebestDNAassemblymethodforyourneeds,pleaseuseourSyntheticBiology/DNAAssemblySelectionChart.
  
  ForhelpdesigningyourprimersforusewithNEBuilder,pleaseviewourprimerdesignvideo.
  
  Figure1:OverviewoftheNEBuilderHiFiDNAAssemblyMethod
  
  Figure2:NEBuilderHiFiDNAAssemblyoffersimprovedefficiencyandaccuracyover
  NEBGibsonAssembly
  Reactionsweresetupina2-and6-fragmentassemblyreactionaccordingtorecommendedreactionconditions.NEBuilderHiFiDNAAssemblyresultsinlargernumbersofcoloniesoverNEBGibsonAssembly,forboth2-and6-fragmentassemblies.
  
  ViewadditionalperformancedatacomparedtoNEBGibsonAssembly
  Figure3:NEBuilderHiFidelivershighercolonyyieldthanIn-FusionHD
  Two-fragmentreactionsweresetupusingthepositivecontrolfromtheIn-FusionHDCloningKit(ClontechTakaraBioUSA,Inc),accordingtorecommendedprotocols.2μlofassemblyreactionwastransformedintosuppliedcompetentcells.1/50ofoutgrowthwasspreadonanAp^Rplate.
  
  ViewadditionalperformancedatacomparedtoIn-FusionHD
  
  ?
  ComparisonofDNAAssemblyReactionTypes
  
  ?
  KitComponents
  Thefollowingreagentsaresuppliedwiththisproduct:
  Storeat(°C) Concentration
  NEBuilder?High-FidelityMasterMix -20
  NEBuilder?PositiveControl -20 2X
  Notes:
  ToensurethesuccessfulassemblyandsubsequenttransformationofassembledDNAs,NEBrecommendsthefollowing:DNA:PCRproductpurificationisnotnecessaryifthetotalvolumeofallPCRproductsis20%orlessoftheassemblyreactionvolume.HighervolumesofPCRproductsmayreducetheefficiencyofhigh-fidelityDNAassemblyandtransformationduetotheelevatedcarryoveramountsofPCRreactionbufferandunusedprimerspresentinthePCRproduct.ColumnpurificationofPCRproductsmayincreasetheefficiencyofbothhigh-fidelityDNAassemblyandtransformationby2–10foldandishighlyrecommendedwhenperformingassembliesofthreeormorePCRfragmentsorassemblinglongerthan5kbfragments.PurifiedDNAforassemblycanbedissolvedinddH2O(Milli-Q?waterorequivalentispreferable),TEorotherdilutionbuffers.Insert:Whendirectlyassemblingfragmentsintoacloningvector,theconcentrationofassemblyfragmentsshouldbeatleast2timeshigherthantheconcentrationofvector.Forassemblyof4ormorefragmentsintoavector,werecommendusinganequimolarratiooffragments.Transformation:NEB5-alphaCompetentE.coli(HighEfficiency,NEB#C2987)providedwiththeNEBuilderHiFiDNAAssemblyCloningKitarerecommendedforuseforassembledproductsoflessthan15kb.ItisalsopossIBLetouseotherNEBcompetentE.colistrains,withtheexceptionofBL21,BL21(DE3),Lemo21(DE3),Nico(DE3),andSHuffle?.WhenusingcompetentE.colifromavendorotherthanNEB,wehavebeendecreasedrobustnessoftransformationwithhigh-fidelityDNAassembledproducts.Electroporation:Electroporationcanincreasetransformationefficiencybyseverallogs.WhenusingtheNEBuilderHiFIDNAAssemblyMasterMix,use1μloftheassembledproductforelectroporation,andplatemultipledilutions.ShouldyourequiretheuseofElectrocompetentcells,pleaseusethe"ElectrocompetentCellsTransformationProtocol"Biology:SomeDNAstructures,includinginvertedandtandemrepeats,areselectedagainstbyE.coli.SomerecombinantproteinsarenotwelltoleratedbyE.coliandcanresultinpoortransformationorsmallcolonies.
售后保障

螞蟻淘生物188，秉承螞蟻淘一貫的嚴謹態度，由螞蟻淘公司專業人員負責品控、采購、物流、銷售、售后，保障正品優質。以“快速好省，為科研提供好產品、好價格”為理念，直接鏈接原廠家，從全國各地原制造商嚴格挑選188款科研精品，剔除品牌溢價，188生物新電商，把好的產品帶給科研！? 力求給你最優質的商品。

Q：生物188產品正品保障嗎？
A：生物188質量把控人員具有十年的從業經驗，在業界享有良好的口碑；自營商城，直接從廠家采購，自己的團隊負責國際物流和清關，中間沒有第三方，所有流程嚴格把控，100%保證正品，假一罰十。

Q：下單后可以修改訂單嗎？
A：下單后的商品付款之前可以修改；訂單付款成功，需要聯系我們客服進行修改；客服電話：4000-520-616

Q：可以開發票嗎？
A：本網站所售商品都是正規清關，均開具16%正規發票，發票金額含配送費金額，另有說明的除外。

Q：商品幾天可以發貨？
A：生物188商品，全部現貨銷售，付款后即可發貨,一般一周內送達！

Q：如何聯系商家？
A：有任何問題夠可以電話咨詢我們，全國24小時免費服務熱線：4000-520-616 或聯系我們的在線客服QQ：1570468124

Q：收到的商品少了/發錯了怎么辦？
A：同個訂單購買多個商品可能會分為一個以上包裹發出，可能不會同時送達，建議查看訂單詳情是否是部分發貨狀態；如未收到，可聯系在線客服或者致電4000-520-616。

Q：退換貨/維修需要多長時間？
A：一般情況下，退貨處理周期為客戶收到產品一個月內（以快遞公司顯示簽收時間為準），包裝規格、數量、品種不符，外觀毀損、短缺或缺陷，請在收到貨24小時內申請退換貨；特殊商品以合同條款為準。

24小時熱銷榜

Parafilm M封口膜/PM996/4x125/盒
￥0.00
Jackson/Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L)/2.0 ml/111-035-003
￥850.00
ThermoFisherQ32856Qubit? Assay Tubes現貨
￥590.00
Qubit? dsDNA BR Assay Kit Q32853現貨
￥2000.00

NEB/NEBuilder? HiFi DNA Assembly Master Mix/E2621S/250 reactions

Description:

KitComponents

Notes:

客戶服務

何為188

關注188微信公眾號

	Storeat(°C)	Concentration
NEBuilder?High-FidelityMasterMix	-20
NEBuilder?PositiveControl	-20	2X