Cambio - Excellence in Molecular Biology

Competent Cells

Competent Cells: Competent cells

A range of electrocompetent and chemically competent cleverly designed E. coli strains for everyday cloning needs through to more demanding applications like CRISPR, working with unstable DNA or low copy number clones. 



TransforMax™ EC100D™ pir plus or pir 116 Electrocompetent E. coli

The TransforMax EC100D pir+ Electrocompetent E. coli and TransforMax EC100D pir-116 Electrocompetent E. coli each express the p protein (the pir gene product) for replication of plasmids containing the R6Kgamma origin of replication (R6Kgamma ori)

BioSearch Tech (Lucigen/Epicentre)

Catalogue No.DescriptionPack SizePriceQty
EC6P095HTransforMax™ EC100D™ pir-116 Electrocompetent E. coli5 X 100µl £319.00 Quantity Add to Order
ECP09500TransforMax™ EC100D™ pir+ Electrocompetent E. coli5 X 100µl £319.00 Quantity Add to Order

Description

The TransforMax™ EC100D™ pir Electrocompetent E. coli and TransforMax™ EC100D™ pir-116 Electrocompetent E. coli each express the p protein (the pir gene product) for replication of plasmids containing the R6Kγ origin of replication (R6Kγ ori). The cells are derived from Epicentre's® TransforMax™ E. coli Electrocompetent E. coli by P1 phage transduction with a strain containing the pir or pir-116 gene linked to a dihydrofolate reductase (DHFR) marker.

The TransforMax™ EC100D™ pir Electrocompetent E. coli will maintain R6Kγori-containing plasmids at approximately 15 copies per cell,1 for cloning of potentially toxic or unstable DNA sequences. The TransforMax™ EC100D™ pir-116 Electrocompetent E. coli are for high copy propagation of up to 250 rescue plasmid copies per cell.1

EZ-Tn5™ Transposons containing the R6Kγori can be introduced into genomic DNA in vivo or plasmid DNA in vitro for 'rescue' cloning applications using the EZ-Tn5™ <R6Kγori /KAN-2> Tnp Transposome or the EZ-Tn5™ <R6Kγori /KAN-2> Insertion Kit, respectively. You can also make your own R6Kγori containing EZ-Tn5™ Transposon using the EZ-Tn5™ pMOD-3 <R6Kγori /MCS> Transposon Construction Vector.

 

Transformation Efficiency

 

Greater than 1X109 colony forming units (cfu)/µg supercoiled DNA

 

Genotypes

 

TransforMax™ EC100D™ pir Electrocompetent E. coli

Maintains plasmids at approximately 250 copies per cell.

F- mcrA delta(mrr-hsdRMS-mcrBC) omega80dlacZdeltaM15 deltalacX74 recA1 endA1 araD139 delta(ara, leu)7697 galU galK γ- rpsL nupG pir-116(DHFR).

 

Important Phenotypes & Applications

 

    • Expresses the <pi> Protein (pir gene product) - for rescue cloning and propagation of vectors containing the R6Kγ origin of replication

 

    • Restriction minus - for efficient cloning of methylated genomic DNA

 

    • Accepts large clones for unbiased propagation and stability of large rescue clones

 

    • Endonuclease minus (endA1) to ensure high yields of plasmid clones

 

    • Recombination minus (recA1) - for stability of large clones

 

  • Compatible with vectors expressing the LacZ™ a-complementing peptide - for blue/white screening of recombinants

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Protocols

Protocols for: TransforMax™ EC100D™ Electrocompetent E. coli

TransforMax™ EC100D™ pir Electrocompetent E. coli & TransforMax™ EC100D™ pir-116 Electrocompetent E. coli Protocol

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References

References

 

  1. 1. Metcalf, W.W. et al. (1994) Gene 138, 1.

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Notes

Note: For transformation with plasmids not having an R6Kγori, use high efficiency TransforMax™ EC100™ Electrocompetent E. coli.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Applications & Benefits

  • Rescue cloning of EZ-Tn5™ transposed elements
  • Rescue transposition mutants created using the EZ-Tn5™ R6KgammaoriKan-2> Insertion Kit
    or Tnp Transposome™ Kit
  • Propagate plasmids tagged with the EZ-Tn5™ R6KgammaoriKan-2> Insertion Kit

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200