當前位置： NEB >>> NEB/Q5??Site-Directed Mutagenesis Kit (Without Competent Cells)/E0552S/10 reactions

NEB/Q5??Site-Directed Mutagenesis Kit (Without Competent Cells)/E0552S/10 reactions

價格: ￥1572.00 市場價: 2620.00

打印產品信息

貨號: E0552S-10reactions

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:
  TheQ5Site-DirectedMutagenesisKit(WithoutCompetentCells)enablesrapid,site-specificmutagenesisofdouble-strandedplasmidDNAinlessthan2hours(Figure1).ThekitutilizestherobustQ5HotStartHigh-FidelityDNAPolymerasealongwithcustommutagenicprimerstocreateinsertions,deletionsandsubstitutionsinawidevarietyofplasmids.AfterPCR,theamplifiedmaterialisaddeddirectlytoauniqueKinase-Ligase-DpnI(KLD)enzymemixforrapid(5minutes),roomtemperaturecircularizationandtemplateremoval(Figure2).Transformationintohigh-efficiencychemically-competentE.coli,notsupplied,ensuresrobustresultswithplasmidsuptoatleast20kbinlength.
  
  Figure1:Site-specificmutagenesisproceedsinlessthan2hours.
  
  Theuseofamastermix,auniquemulti-enzymeKLDenzymemix,andafastpolymeraseensuresthat,formost?plasmids,themutagenesisreactioniscompleteinlessthantwohours.
  ???
  Figure2:Q5Site-DirectedMutagenesisOverview.
  
  Thiskitisdesignedforrapidandefficientincorporationofinsertions,deletionsandsubstitutionsintodoublestranded?plasmidDNA.Thefirststepisanexponentialamplificationusingstandardprimersandamastermix?fomulationofQ5HotStartHigh-FidelityDNAPolymerase.Thesecondstepinvolvesincubationwithaunique?enzymemixcontainingakinase,aligaseandDpnI.Together,theseenzymesallowforrapidcircularizationofthe?PCRproductandremovalofthetemplateDNA.Thelaststepisahigh-efficiencytransformationintochemicallycompetent?cells(notprovided).
  Figure3:PrimerDesignforQ5Site-DirectedMutagenesis
  
  Substitutions,deletionsandinsertionsareincorporatedintoplasmidDNAthroughtheuseofspecificallydesigned?forward(black)andreverse(red)primers.Unlikekitsthatrelyonlinearamplification,primersdesigned?fortheQ5Site-DirectedMutagenesisKitshouldnotoverlaptoensurethatthebenefitsof?exponentialamplificationarerealized.A)Substitutionsarecreatedbyincorporatingthedesirednucleotide?change(s)(denotedby*)inthecenteroftheforwardprimer,includingatleast10complementarynucleotideson?the3′sideofthemutation(s).Thereverseprimerisdesignedsothatthe5′endsofthetwoprimersannealbackto-back.B)Deletionsareengineeredbydesigningstandard,non-mutagenicforwardandreverseprimersthatflank?theregiontobedeleted.C)Insertionslessthanorequalto6nucleotidesareincorporatedintothe5′endofthe?forwardprimerwhilethereverseprimerannealsback-to-backwiththe5′endofthecomplementaryregionofthe?forwardprimer.D)Largerinsertionscanbecreatedbyincorporatinghalfofthedesiredinsertionintothe5′ends?ofbothprimers.Themaximumsizeoftheinsertionislargelydictatedbyoligonucleotidesynthesislimitations.
  Figure4:NEB’sQ5SDMKitdelivershighertransformationefficiencythanAgilent’sQuikChange^?SDMKit
  Resultsfromasubstitutionreaction(4nt)usingtheback-to-backControlSDMPrimerMixandControlSDMPlasmid(6.7kb)areshown,alongwithresultsfroma12ntdeletionexperiment(5.8kbplasmid)andan18ntinsertionexperiment(7.0kbplasmid).Inallthreecases,over90%oftheresultantcolonieshadincorporatedthedesiredmutation(s).Resultsarenormalizedtototaltransformantsifcellswerenotdilutedpriortoplating.Forcomparison,thesamesubstitutionreaction(4nt)wasperformedwiththeQuikChangeLightningSite-DirectedMutagenesisKit(Agilent)followingAgilent’sprotocolandusingAgilent’sprimerdesigntooltodesignoverlappingprimers.
  
  *NotethattheQuikChangekitdoesnotaccommodatedeletionsandinsertionsofthissize,sonocomparisoncouldbemadefortheseexperiments.
  KitComponents
  Thefollowingreagentsaresuppliedwiththisproduct:
  Storeat(°C) Concentration
  Q5®HotStartHigh-Fidelity2XMasterMix -20 2X
  KLDEnzymeMix -20 10X
  KLDReactionBuffer -20 2X
  ControlSDMPrimerMix -20 10μM
  ControlSDMPlasmid -20 5μg/ml
  Notes:
  StorageNote:TheQ5Site-DirectedMutagenesisKit(WithoutCompetentCells)isstableat–20°Cfortwoyears.Forflexibility,themutagenesisreagentsandcontrolreactionsaresuppliedwithoutcompetentcellssothatanychemically-competentE.colicellssuitableforcloningmaybeused.
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/Q5??Site-Directed Mutagenesis Kit (Without Competent Cells)/E0552S/10 reactions

Description:

KitComponents

Notes:

客戶服務

何為188

關注188微信公眾號

	Storeat(°C)	Concentration
Q5®HotStartHigh-Fidelity2XMasterMix	-20	2X
KLDEnzymeMix	-20	10X
KLDReactionBuffer	-20	2X
ControlSDMPrimerMix	-20	10μM
ControlSDMPlasmid	-20	5μg/ml