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蚂蚁淘在线 / 品牌中心 / Lucigen / Lucigen/EZ-Tn5™ <KAN-2>Tnp Transposome™ Kit/TSM99K2/10 Reactions

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

价格
￥10000.00
货号：TSM99K2
浏览量：127
品牌：Lucigen
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商品描述

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. Lucigen基因组编辑和工程面临许多挑战。为您的编辑实验获取可靠的试剂不应该是其中之一。无论您是在突破CRISPR技术的界限，在体内产生基因敲除或敲入还是在进行序列消化或鉴定的体外反应，您都需要可靠的工具和酶。依靠CRISPRcraft™获得质量，可靠性和稳定的性能。