當前位置： NEB >>> NEB/SplintR? Ligase/M0375S/1,250 units

NEB/SplintR? Ligase/M0375S/1,250 units

價格: ￥1092.00 市場價: 1820.00

打印產品信息

貨號: M0375S-1,250units

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:
  SplintRLigase,alsoknownasPBCV-1DNALigaseorChlorellavirusDNALigase,efficientlycatalyzestheligationofadjacent,single-strandedDNAsplintedbyacomplementaryRNAstrand.ThispreviouslyunreportedactivitymayenablenovelapproachesforcharacterizationofmiRNAsandmRNAs,includingSNPs.SplintRisideallysuitedformanytargetenrichmentworkflowswithapplicationsinnext-generationsequencingandmoleculardiagnostics.TherobustactivityoftheenzymeanditsaffinityforRNA-splintedDNAsubstrates(apparentKm=1nM)enablesub-nanomolardetectionofuniqueRNAspecieswithinacomplexmixture,makingSplintRligaseasuperiorchoicefordemandingRNAdetectiontechnologies.
  
  TheenzymeisactiveoverabroadrangeofATPconcentrations(10μM–1mM)andpH(6.5–9).OptimalactivityisobservedusingMg²⁺>5mM,andapHbetween7.5and8.0.Theactivityisenhancedathighertemperatures(upto37°C),andbysupplementationwith5mMMn²⁺.Thereactionisinhibitedbysaltconcentrationsinexcessof100mM.
  
  Theenzymetoleratesallbasepaircombinationsattheligationjunction,butispartiallyinhibitedbydC/GanddG/Cbasepairsatthedonor(phosphorylated)sideligationjunction,particularlywhenthe+2basewasalsoaC/Gbasepair.
  
  LigationofDNAsplintedbyRNA
  
  (A)Outlineoftheligationassay:a5′-phosphorylated,3′-FAMlabeledDNA“donor”oligonucleotideandanunmodifiedDNA“acceptor”oligonucleotideare?annealedtoacomplementaryRNAsplint.Thissubstrateisreactedwithaligasetoformamixtureofunreactedstartingmaterial(I),adenylylatedDNA(II),andligatedproduct(III).These?productsaredenatured,separatedbycapillaryelectrophoresisanddetectedbyfluorescence.(B)LigationoftheRNA-splintedsubstrateinSplintRLigaseReactionBufferfor15minutesat?25°Cwith(a)noenzyme,(b)1μMT4DNALigaseand(c)100nMSplintRLigase.IndicatedpeakscorrespondtostartingpDNA(I),AppDNA(II)andligatedproduct(III)asdeterminedby?co-elutionwithsyntheticallypreparedstandards.(C)ThefractionofligatedproductcatalyzedbyeitherSplintRLigaseorT4DNALigasewasanalyzedbyperformingsetsofligationswith?bothligasesatconcentrationsbetween10pMand10μMfor15minutesat25°C.SplintRLigaseisclearlymuchmoreefficientatligationofRNAsplintedDNAthanT4DNALigase.
  ProductSource
  AnE.colistrainthatcarriesarecombinantgeneencodingPBCV-1DNALigase.
  ReagentsSupplied
  Thefollowingreagentsaresuppliedwiththisproduct:
  Storeat(°C) Concentration
  SplintRLigaseReactionBuffer 10X
  Notes:
  SplintRLigaseisinhibitedbymonovalentcations.Westronglyrecommendensuringthesecommonreactants(NaCl,KCl)arekepttobelow50mMinthereaction.Theenzymeissuppliedinastoragebuffercontaining300mMNaCl,forstoragestABIlity.Aminimum6-folddilutionoftheenzymebyadditiontothereactionisrecommended,withtheoptimaldilutionbeing≥10-fold.Ifdilutionofenzymeforstorageisneeded,werecommendusingDiluentA(NEB#B8001).ReactionswithSplintRshouldbeperformedbetween16–37°C.Werecommendinitialtestingbeperformedat25°C.Theenzymeissuppliedasa10.5μMsolution.Wesuggestmaintainingtheenzymebelow1μMinthereactionwithasuggestedrangeof100nMto1μM.Formanyapplications,startingwitha2-foldexcessofenzymeoverligatableendsisideal.Forexample,intheexperimentdescribedintheaccompanyingfigureasubstrateconcentrationof100nMwasfoundtobeideal.Inthatworkflow,250nMenzymegavecompleteligationin15minutesonallsequencestested.Ifthereactionisnotproceedingasefficientlyasdesired,westronglyrecommendextendingtheincubationtimeratherthanincreasingtheconcentrationofenzymeinthereactionbeyond1μM.AnalternativeoptionforrecalcitrantsubstratesistousealowconcentrationofATP.AlowATPbuffercangivehigheryieldsofligationproductforsubstratesthathavelowligationefficienciesinthestandardSplintRLigaseReactionBuffer,suchassubstrateswithrunsofG:Cbasepairsattheligationjunction.Wesuggest1XT4RNALigaseReactionBuffer(NEB#B0216)supplementedwithATP(NEB#P0756)toafinalconcentrationof10μM.WerecommendanRNAsplintofatleast20complementarybases.Wehavefound10bpofdsDNA/RNAoneithersideofthejunctiontobesufficientforallsubstratestested.Thesplintdoesnothavetobecenteredontheligationjunction,however,withasfewasfourbasesononesideofthejunctiongivingcompleteligationforasplintwith20basesoftotalcomplementarity,dependingonsubstratesequence.Ifregionsofoverlap<10bparedesired,sometestingwillberequiredtodeterminetheminimumlengthofthedsregionforyourspecificsequence.InordertomaximizefidelityinligationsbySplintRDNALigase,pleaseconsiderthefollowing:IncreasingreactiontemperaturehasbeenshowntoimproveactivityandspecificityofSplintRDNALigase.Incubationat37°CgivesimprovedresultsindetectionofmicroRNA[2].AdditionofETSSB(NEB#M2401)hasalsobeenshowntoincreaseactivityandreduceoff-targetligationwhenligatingDNAprobestoRNAusingSplintRDNALigase[3].
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/SplintR? Ligase/M0375S/1,250 units

Description:

ProductSource

ReagentsSupplied

Notes:

客戶服務

何為188

關注188微信公眾號

	Storeat(°C)	Concentration
SplintRLigaseReactionBuffer		10X