Lucigen/TransforMax™ EPI300™ Electrocompetent E. coli/EC300110/10 X 100 µl

StandardcloningforlargeinsertsandGibsonassemblyelectroporation

Clonelargeinsertsandtransformlargeplasmids-uptoatleast145kbplasmidDNA.
GeneratecloneswithinduciblecopynumberusingCopyControl™vectors.
Achievehightransformationefficiencies:>1×10¹⁰cfu/µgpUC19DNA.
IncreaseGibsonAssembly®colonycountswithhigh-efficiencyelectroporation

FreeSampleAvailable

Applications

Generationofinduciblecopy-numberclonesusingtheCopyControl™CloningSystem.

TransforMax™EPI300™E.colicellslackthetonAgeneandareengineeredforusewithEpicentre'sCopyControl™cDNA,Gene,andPCRCloningKitandotherCopyControlCloningSystems*thatdonotrequirephageT1-resistantcells.ThecellscontainaninduciblemutanttrfAgenewhosegeneproductisrequiredforinitiationofreplicationfromtheoriVoriginofreplication,suchasthatinCopyControlpCC1™vectors.OnLBchloramphenicolplatesorinLBorSOCmediasupplementedwithchloramphenicol,CopyControlclonesgrowninTransforMaxEPI300E.colireplicateatsingle-copynumberfromtheF-factorrepliconbecauseexpressionofthetrfAgeneisrepressed.AdditionofCopyControlInductionSolutioninducesthecellstoexpressthetrfAgeneproductandinducestheirreplicationathigh-copynumberfromoriV(Fig.1).ReplicationfromoriVresultsinhigheryieldsandhigherpurityofclonedDNA.

Benefits

Copy-numberofclonesisundertightcontrolofaninduciblepromoterlinkedtothetrfAgene.
Hightransformationefficiencywithclonesofallsizes.
lacZΔM15forblue/whitescreeningofrecombinants.
Restriction-minus[mcrA,Δ(mrr-hsdRMS-mcrBC)]phenotypeenablesefficientcloningofmethylatedDNA.
Endonuclease-minus(endA1),toensurehighyieldsofDNA.
Recombination-minus(recA1),forgreaterstabilityoflargeinsertclones.

Figure1.CopynumberofCopyControl™BACclonescanbeinduced10-to20-foldinTransforMax™EPI300™E.coli.TheyieldofBACDNAfromCopyControlBACclonesof110-145kbincreased>14-foldfollowingadditionofCopyControlInductionSolution.U=uninducedcells;I=inducedcells.

Genotype

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

TransforMaxEPI300ElectrocompetentE.coli

Transformationefficiencyof>1x10¹⁰cfu/µgofpUC19.

TransforMaxEPI300ChemicallyCompetentE.coli

Transformationefficiencyof>5x10⁸cfu/µgofpUC19.

*Coveredbyissuedand/orpendingpatents.

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1,000Xconcentratedsolution.Filtersterilized.

对照品标准品，生物试剂盒，gibco胎牛血清，试剂采购网，中国试剂网，细胞因子试剂盒

Lucigen/TransforMax™ EPI300™ Electrocompetent E. coli/EC300110/10 X 100 µl

StandardcloningforlargeinsertsandGibsonassemblyelectroporation

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