Applications

• Constructionofinducible-copy-numbergenomiclibrariesusingtheCopyControl™CloningSystem,withclonesthatareresistanttocontaminatingphageT1andT5.

PhageT1-ResistantTransforMax™EPI300™-T1^RE.colihaveallthebenefitsofthehighlyversatileTransforMaxEPI300™E.colicompetentcellswiththeadditionofbeingresistanttobacteriophagesT1andT5(tonAgenotype).Onceintroducedintothelabenvironment,bacteriophageT1rapidlylysesE.colistrainsthatarecommonlyusedincloningapplicationsandresultsinsignificantlabdowntimeandthelossofvaluableclones.BacteriophageT1isparticularlydifficulttoeliminatefromthelabandcanlaydormantformanyyears.ThetonAgenotypeprotectsthephageT1-resistantTransforMaxEC100-T1^RcellsandvaluableclonesfromattackbybacteriophageT1.

LikethestandardTransforMaxEPI300E.coli,thisstrainhasbeenspeciallyengineeredforusewithEpicentre'sCopyControl™CloningSystems.*ThecellscontainaninduciblemutanttrfAgenewhosegeneproductisrequiredforinitiationofreplicationfromtheoriVcontainedontheCopyControlpCC1™VectorsoronclonesthathavebeenretrofittedwithCopyControlcapABIlityusingtheEZ-Tn5™<oriV/KAN-2>InsertionKit.ThisprocessallowsTransforMaxEPI300-T1^RE.colitomaintainCopyControlvectorsatsinglecopyuntilinductionoccursimmediatelybeforeDNApurification.

Benefits

• tonAforresistancetobacteriophagesT1andT5.
• trfAgeneundertightcontrolofaninduciblepromoterforcopy-numbercontrolofCopyControlclonesandclonesretrofittedwiththeEZ-Tn5<oriV/KAN-2>Transposon.
• Hightransformationefficiencyofbothlargeandsmallclones.
• lacZΔM15forblue/whitescreeningofrecombinants.
• ReADIlyacceptslargeDNAsforconstructionoflarge-insertgenomiclibraries.
• Restrictionminus[mcrA,Δ(mrr-hsdRMS-mcrBC)]enablesefficientcloningofmethylatedDNA.
• Endonucleaseminus(endA1)toensurehighyieldsofDNA.
• Recombinationminus(recA1)forgreaterstabilityoflargeclonedinserts.

Genotype

F^-mcrAΔ(mrr-hsdRMS-mcrBC)Φ80dlacZΔM15ΔlacX74recA1endA1araD139Δ(ara,leu)7697galUgalKλ^-rpsL(Str^R)nupGtrfAtonA

*Coveredbyissuedand/orpendingpatents.