StandardcloningforlargeinsertsandGibsonassemblyelectroporation
- Clonelargeinsertsandtransformlargeplasmids-uptoatleast145kbplasmidDNA.
- GeneratecloneswithinducIBLecopynumberusingCopyControl™vectors.
- Achievehightransformationefficiencies:>1×1010cfu/µgpUC19DNA.
- IncreaseGibsonAssembly®colonycountswithhigh-efficiencyelectroporation
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
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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(StrR)nupGtrfAdhfr
TransforMaxEPI300ElectrocompetentE.coli
- Transformationefficiencyof>1x1010cfu/µgofpUC19.
TransforMaxEPI300ChemicallyCompetentE.coli
- Transformationefficiencyof>5x108cfu/µgofpUC19.
*Coveredbyissuedand/orpendingpatents.