Stabilize toxic targets in favorite cloning and expression vectors.
- Stabilize toxic inserts in common cloning and expression vectors (pUC and pET-type vectors)
- Clone and maintain challenging sequences at reduced plasmid copy number, then induce to high copy number for DNA recovery
- Avoid T1 and T5 phage contamination with tonA mutation
- Choose chemically competent cells for general cloning or electrocompetent cells for demanding applications such as library generation
Simplify your cloning strategy for unstable DNA, and get to the next phase of your project faster.
DNA that is unstable at high-copy number often codes for a protein that inhibits cell growth or contains AT- and GC-rich sequences or sequences with strong secondary structure. Lowering the plasmid copy number is a common strategy to successfully clone unstable DNA, but often requires use of an alternate vector engineered with a low-copy origin of replication. CopyCutter™ EPI400™ E. coli cells are engineered to significantly lower the copy number of a wide variety of common vectors, so you can clone unstable DNA sequences using your favorite cloning and expression vectors.
The CopyCutter EPI400 cell line was created 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 parental strain and replaced with a modified pcnB gene 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 strain (a derivative of EC100) is approximately 4- to 25-fold lower, depending on the vector. Increase copy number to improve plasmid yields after a short incubation with the CopyCutter Induction Solution.
CopyCutter competent cells are recombination minus (recA) and endonuclease minus (endA), for high yield, high quality plasmid DNA preparations. The strain is also blue/white screening capable (lacZΔM15) and restriction minus [mcrA, Δ(mrr-hsdRMS-mcrBC)] for cloning methylated DNA.
CopyCutter EPI400 Electrocompetent E. coli
Transformation efficiency of >1 x 1010 cfu/µg of pUC19
CopyCutter EPI400 Chemically Competent E. coli
Transformation efficiency of >1 x 107 cfu/µg of pUC19
Delivery information: Each kit contains ten tubes with 50 µL of cells (enough cells for 10 transformations), CopyCutter™ Induction Solution and pUC19 control DNA. The CopyCutter™ Induction Solution is provided at a 1000X concentration and is filter sterilized.
Caution: For research use only. Not for human or diagnostic use.
