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卢西根/TransforMax™EC100D™pir-116电子兼容大肠杆菌/EC6P095H/5 x 100µl
RescuecloningofEZ-Tn5™transposedelements
- RescuetranspositionmutantscreatedusingtheEZ-Tn5™R6KɣoriKan-2>InsertionKit
orTnpTransposome™Kit - 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
- Metcalf,W.W.etal.(1994)Gene138,1.