Description
Perform plasmid rescue for transposition mutants created with the EZ-Tn5 Transposome Kits.
Each product contains 5 tubes each containing 100 µL of electrocompetent cells, which is enough cells for 10 electroporations. A control vector containing and R6Kγori is also included with each product.
Key features
- Rescue cloning of EZ-Tn5 transposed elements
- Propagate plasmids tagged with the EZ-Tn5 Insertion Kit
Product information
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 (R6Kγ ori). 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 R6Kγ ori 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 (Str R) 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 (Str R) nupG pir-116(DHFR)
Transformation Efficiency
Greater than 5 × 10 9 colony forming units (cfu)/µg with pR6Kan control DNA.
MANUAL