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当前位置: 首页 > 产品中心 > In Vitro Tn5 Transposon Mutagenesis > Lucigen/EZ-Tn5™ <R6Kyori/KAN-2> Insertion Kit/EZI011RK/10 Reactions
公司介绍
美国Lucigen公司,自1998年成立至今,一直致力于生命科学领域相关科研产品的研究与开发,在分子生物学领域处于领导性地位。Lucigen公司主要开发各类用于基因克隆的试剂盒及相关产品,包括:CloneSmart®平端克隆试剂盒、BigEasy®线性克隆系统、pEZSeq™平端克隆试剂盒、ClonePlex™ AK文库构建试剂盒、DNA Terminator ®末端修复试剂盒、EconoTaq® DNA聚合酶及E.cloni感受态细胞等
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Lucigen/EZ-Tn5™ <R6Kyori/KAN-2> Insertion Kit/EZI011RK/10 Reactions

Lucigen/EZ-Tn5™ <R6Kyori/KAN-2> Insertion Kit/EZI011RK/10 Reactions
  • Lucigen/EZ-Tn5™ <R6Kyori/KAN-2> Insertion Kit/EZI011RK/10 Reactions
商品介绍

RandominsertionofR6KoriginforrescueofflankingDNAorplasmids

  • InsertR6KoriginrandomlyintoanyDNAsequenceinvitroFormerly from Epicentre
  • Skipprimerwalking-simplifySangersequencingoflargeDNAinserts
  • RescueclonesinE.colihostexpressingthepirgeneproduct
  • MinimizeinsertionbiaswiththehyperactiveTn5system,knownforhighestlevelofrandomness

Applications

  • "Rescue"ofplasmidsoranyothercircularizedDNA(e.g.,mitochondrialDNA)thatwouldnototherwisereplicateinE.colibecausetheylackarecognizableoriginofreplicationand/oraselectableMarker.
  • PreparationofDNAsequencingtemplatesfromtransposoninsertioncloneswithoutprimerwalkingoradditionalsubcloning.
  • Creationofalibraryofrandomgeneknockoutsinvitrotofacilitategeneticanalysisofplasmid-encodedgenes.

TheEZ-Tn5™<R6Kγori/KAN-2>InsertionKit*facilitatesthesequencingandgeneticanalysisofplasmidsoranyothercircularizedDNAthatwouldnototherwisereplicateinE.coli.1,2ThekitisbasedupontheEZ-Tn5™<R6Kγori/KAN-2>TransposonwhichcarriestheE.coliR6Kγconditionaloriginofreplication(R6Kγori)andakanamycinresistancemarker.Asimple,one-step,2-hourinvitroreactionrandomlyinsertsthetransposonintothetargetDNA.ThenanaliquotofthereactionisusedtotransformanE.colihostexpressingthepirgeneproduct,whichisrequiredforreplicationfromtheR6Kγori.Insertionclonesareselectedonkanamycinplatesandcanbesequencedbidirectionallyusingtheprovidedprimersthatarehomologoustotheendsofthetransposon.ClonescanbemaintainedinEpicentre'sTransforMax™EC100D™pir+orTransforMax™EC100D™pir-116strains.3

g_eztn5_r6kgori_kan2_insertionFigure1.AplasmidcontainingtheEZ-Tn5™<R6Kγori/KAN-2>TransposoncanbemaintainedinTransforMax™EC100D™pir+cellsat~15copiespercell(Lanes1-4)orTransforMax™EC100D™pir-116cellsat~250copiespercell(Lanes5-8).ColoniesfromrandomlychosencloneswereprocessedusingtheColonyFast-Screen™Kit.A5-microliteraliquotoflysatewasloadedperlane.M,supercoiledDNAladder.

References

  1. Jendrisak,J.etal.(2002)EpicentreForum9(1),14.
  2. Yoon,Y.andKoob,M.(2003)EpicentreForum10(2),10.
  3. Metcalf,W.W.etal.(1994)Gene138,1.

*Coveredbyissuedand/orpendingpatents,exclusivelylicensedorassignedtoEpicentre®(anIllumina®Company).

ORDERINFORMATION

EZ-Tn5™Transposase,EZ-Tn5™<R6Kyori/KAN-2>Transposon,EZ-Tn5™10XReactionBuffer,EZ-Tn5™10XStopSolution,ForwardandReversePrimers,ControlTargetDNA,SterileWater.
品牌介绍

美国Lucigen公司,自1998年成立至今,一直致力于生命科学领域相关科研产品的研究与开发,在分子生物学领域处于领导性地位。Lucigen公司主要开发各类用于基因克隆的试剂盒及相关产品,包括:CloneSmart®平端克隆试剂盒、BigEasy®线性克隆系统、pEZSeq™平端克隆试剂盒、ClonePlex™ AK文库构建试剂盒、DNA Terminator ®末端修复试剂盒、EconoTaq® DNA聚合酶及E.cloni感受态细胞等。Lucigen公司凭借其独到的产品技术,过硬的产品质量,良好的产品服务赢得了全球广大用户的信赖。

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