Description
General description
CopyCutter EPI400 E.coli cells were developed to stabilize toxic inserts and cloning toxic unstable DNA.
CopyCutter EPI400 E.coli cells significantly lower the copy number of a wide variety of popular high copy number vectors to maintain “toxic” genes or unstable DNA sequences in the vectors.
The CopyCutter EPI400 E.coli Induction Solution is provided to induce to higher copy number for improved plasmid yield.
The CopyCutter EPI400 E.coli cell line was derived from Epicentre′s high-transformation efficiency phage T1-resistant TransforMAX EC100-T1R E.coli strain by manipulating a gene that controls the copy number of vectors containing ColE1 or pMB1 origins of replication (e.g., pUC- and pET-type vectors). This constitutively expressed gene, pcnB (plasmid copy number), was deleted from the TransforMAX EC100 strain and replaced with a modified pcnB gene that is linked to an inducible promoter, creating the CopyCutter EPI400 strain.
The copy number of ColE1-type vectors in the CopyCutter EPI400 strain compared to the parental TransforMAX EC100 strain is approximately 4- to 25-fold lower, depending on the vector.
1 transformation per tube, 10x50µl.
Application
Cloning of unstable DNA sequences;
Cloning of expressing toxic proteins
Features and Benefits
Optimized for cloning toxic or unstable DNA
Stabilize toxic inserts in common cloning and expression vectors (pUC and pET-type vectors)
Maintain clones at a low-copy number, then induce to higher copy number for improved plasmid yield
Avoid T1 and T5 phage contamination with tonA mutation
Choose chemically competent cells (LGCC400CH10) for general cloning or electrocompetent cells (LGCC400EL10) for demanding applications such as library generation
Transformation efficiency: ≥ 1 x 1010 (cfu/µg pUC DNA)
Genotype: CopyCutter EPI400: F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL (StrR) nupG trfA tonA pcnB4 dhfr
Components:
CopyCutter EPI400 ElectroCompetent Cells
pUC 19 Trasformation Control
CopyControl Induction Solution
