TransforMax™ EC100D™ pir+ and pir-116 Electrocompetent E. coli
Applications
- Rescue cloning of transposition mutants created using the EZ-Tn5™ <R6Kγori/KAN-2> Insertion kit or Transposome™, or the HyperMu™ <R6Kγori/KAN-1>Tnp Transposome.
- Propagation of plasmids tagged with the EZ-Tn5 <R6Kγori/KAN-2> Insertion kit.
The TransforMax™ EC100D™ pir+ Electrocompetent E. coli and TransforMax™ EC100D™ pir-116 Electrocompetent E. coli each constitutively express the π protein (the pir gene product) for replication of plasmids containing the R6Kγ origin of replication (R6Kγori). The cells are derived from Epicentre’s TransforMax™ EC100™ 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+ cells 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 cells are for high-copy propagation of up to 250 rescue plasmid copies per cell.1
Benefits
- Accepts large clones for unbiased propagation and stability of large rescue clones.
- lacZΔM15 for blue/white screening of recombinants.
- Restriction minus [mcrA, Δ(mrr-hsdRMS-mcrBC)] enables efficient cloning of methylated DNA.
- Endonuclease minus (endA1) to ensure high yields of DNA.
- Recombination minus (recA1) for greater stability of large cloned inserts.
Genotypes
TransforMax EC100D pir+ Electrocompetent E. coli:
F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL (StrR) nupG pir+(DHFR)
TransforMax EC100D pir-116 Electrocompetent E. coli:
F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL (StrR) nupG pir-116(DHFR)
Transformation Efficiency
Greater than 1 x 109 colony forming units (cfu)/µg with pR6Kan control DNA.
References
- Metcalf, W.W. et al. (1994) Gene 138, 1.
Ordering
Catalog No. | Size |
---|
TransforMax™ EC100D™ pir+ Electrocompetent E.coli
ECP09500 | 5 x 100 µl |
TransforMax™ EC100D™ pir-116 Electrocompetent E.coli
EC6P095H | 5 x 100 µl |