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Lucigen/TransforMax™ EC100D™ pir+ Electrocompetent E.coli/ECP09500/5 x 100 µl

RescuecloningofEZ-Tn5™transposedelements

  • RescuetranspositionmutantscreatedusingtheEZ-Tn5™R6KɣoriKan-2>InsertionKit
    orTnpTransposome™KitFormerly from Epicentre
  • PropagateplasmidstaggedwiththeEZ-Tn5™R6KɣoriKan-2>InsertionKit

TheTransforMax™EC100D™pir+ElectrocompetentE.coliandTransforMax™EC100D™pir-116ElectrocompetentE.colieachconstitutivelyexpresstheπprotein(thepirgeneproduct)forreplicationofplasmidscontainingtheR6Kγoriginofreplication(R6Kγori).ThecellsarederivedfromEpicentre'sTransforMax™EC100™ElectrocompetentE.colibyP1phagetransductionwithastraincontainingthepir+orpir-116genelinkedtoadihydrofolatereductase(DHFR)Marker.

TheTransforMaxEC100Dpir+cellswillmaintainR6Kγoricontainingplasmidsatapproximately15copiespercell,1forcloningofpotentiallytoxicorunstableDNAsequences.TheTransforMaxEC100Dpir-116cellsareforhigh-copypropagationofupto250rescueplasmidcopiespercell.1

Benefits

  • AcceptslargeclonesforunbiasedpropagationandstABIlityoflargerescueclones.
  • lacZΔM15forblue/whitescreeningofrecombinants.
  • Restrictionminus[mcrA,Δ(mrr-hsdRMS-mcrBC)]enablesefficientcloningofmethylatedDNA.
  • Endonucleaseminus(endA1)toensurehighyieldsofDNA.
  • Recombinationminus(recA1)forgreaterstabilityoflargeclonedinserts.

Genotypes

TransforMaxEC100Dpir+ElectrocompetentE.coli:
F-mcrAΔ(mrr-hsdRMS-mcrBC)φ80dlacZΔM15ΔlacX74recA1endA1araD139Δ(ara,leu)7697galUgalKλ-rpsL(StrR)nupGpir+(DHFR)

TransforMaxEC100Dpir-116ElectrocompetentE.coli:
F-mcrAΔ(mrr-hsdRMS-mcrBC)φ80dlacZΔM15ΔlacX74recA1endA1araD139Δ(ara,leu)7697galUgalKλ-rpsL(StrR)nupGpir-116(DHFR)

TransformationEfficiency
Greaterthan5×109colonyformingunits(cfu)/µgwithpR6KancontrolDNA.

References

  1. Metcalf,W.W.etal.(1994)Gene138,1.

ORDERINFORMATION

Eachproductcontains5tubeseachcontaining100µLofelectrocompetentcells,whichisenoughcellsfor10electroporations.AcontrolvectorcontainingandR6Kγoriisalsoincludedwitheachproduct.
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