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当前位置: 首页 > 产品中心 > In Vivo Tn5 Transposon Mutagenesis > Lucigen/EZ-Tn5™ <KAN-2>Tnp Transposome™ Kit/TSM99K2/10 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™ <KAN-2>Tnp Transposome™ Kit/TSM99K2/10 Reactions

Lucigen/EZ-Tn5™ <KAN-2>Tnp Transposome™ Kit/TSM99K2/10 Reactions
  • Lucigen/EZ-Tn5™ <KAN-2>Tnp Transposome™ Kit/TSM99K2/10 Reactions
商品介绍

Generaterandomgeneknockoutsinlivingbacteria

  • GeneratemutantswithimprovedgeneticsorfunctionacrossabroadhostrangeofbacterialcelltypesFormerly from Epicentre
  • Characterizenovelgenesandgenefunctions
  • Identifygenesinvolvedinpathogenesis,toxicity,biofilmdevelopment
  • Unravelmetabolicpathways
  • Identifyessentialgenesandregulatoryelements
  • 100’sofcitationsformanydifferentapplications

Applications

  • Rapidgenerationofknock-outmutantsinbacterialcells.
  • Knock-inofgenesforbacterialstraindevelopment.
  • "Tagging"bacteriawithvisIBLegeneticMarkersforenvironmentallocalizationstudies.
  • DirectsequencingofbacterialchromosomalDNA.

EZ-Tn5™Transposome™complexesareformedbetweenanEZ-Tn5™TransposonandEZ-Tn5™Transposase,andprovideasimpleandreliablemethodforgeneratingalibraryofrandomgeneknockoutsinvivo.*JustelectroporatetheEZ-Tn5TransposomeintoanyofabroadrangeoflivingbacterialcellsandselectforamarkerencodedbytheEZ-Tn5Transposon(Fig.1).Becausethereisnoneedforcellconjugation,suicidevectors,orspecifichostfactors,EZ-Tn5Transposomesareidealforcreatingmutantsinspeciesthathavepoorlydescribedgeneticsystemsorlackadequatemoleculartools.

Ready-to-useEZ-Tn5Transposomes*areavailablecontainingakanamycinselectablemarker(<KAN-2>).ThismarkerisreADIlyexpressedinmanyGram-negativebacteria.YoucanalsocreateyourownEZ-Tn5TransposomeusingoneoftheEZ-Tn5pMOD™TransposonConstructionVectorsandEZ-Tn5Transposase.

Figure1.TheEZ-Tn5™TransposoninsertionsiteinbacterialDNAcanbesequenceddirectlyusinggenomicDNAisolatedusingtheMasterPure™CompleteDNAPurificationKitandprimershomologoustotheendsofthetransposon.

AllEZ-Tn5Transposonscontainuniqueprimer-bindingsitesateachendforbidirectionalsequencing.Hence,ageneknockoutcanbesequenceddirectlyusingbacterialgenomicDNAastemplateandtheprimersprovidedwitheachTransposome.TransposoninsertionsmadeusinganEZ-Tn5<R6Kγori/KAN-2>TnpTransposomeKitcanberescuedandtheflankingDNAsequenced.

EZ-Tn5Transposome-mediatedinsertionshavebeenmadeinmanydifferentmicroorganisms,includingAcinetobactor,Campylobacter,Escherichia,Mycobacterium,Proteus,Pseudomonas,Saccharomyces,Salmonella,Trypanosoma,Xylella,andothers.Thenumberoftranspositionclonesobtainedishighlydependentonthetransformationefficiencyofthehostcell(Table1).

Benefits

  • Rapidmutagenesisprocedureissimplerandeasiertousethanchemicalmutagenesis.
  • Moreefficientthanusingmini-transposonswithsuicideplasmids.
  • Broadhostrange:over60speciesofGram-negativeandGram-positivebacteriatransposedsofar.
Table1.ExamplesofbacterialstrainsmutagenizedusingtheEZ-Tn5™Tranposomes™
Actinobacilluspleuropneumoniae
Agrobacteriumtumefaciens
Bacillussubtilis
Bartonellahenselae
BDellovibriobacteriovorus
Campylobacterjejuni
Clavibactermichiganensis
subsp.sepedonicus
CoryNEBacteriumdiphtheriae
Enterobactercloacae
Escherichiacoli
Francisellatularensis
Haemophilusducreyi
Moraxellacatarrhalis
Mycobacteriumavium
Mycobacteriumbovis
(BCG)
Mycobacteriumtuberculosis
Myxobacteriumangiococcus
Neisseriagonorrhoeae
Pseudomonasputida
Pseudomonassyringae
Rhodococcusequi
Rickettsiaprowazekii
Salmonellatyphimurium
Serratiamarcesens
Silicibacterpomeroyi
Spiroplasmacitri
Streptococcuspyogenes
Xanthomonascampestris
Xylellafastidiosa
Zymomonasmobilis
Table2.NumberofKanRtransposoninsertionclonesproducedfromelectroporationof1µlofEZ-Tn5™<KAN>TnpTransposome™
E.coli>105Proteusvulgaris>103
Salmonellaty.>104Mycobacteriumsmegmatis>102
Pseudomonassp.>102

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

ORDERINFORMATION

EZ-Tn5™TnpTransposome™,KAN-2FP-1ForwardPrimer,KAN-2RP-1ReversePrimer,andSterileWater.
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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