Cambio - Excellence in Molecular Biology

Molecular Cloning Kits

Molecular Cloning Kits: Competent Cells

CopyCutter™ EPI400™ Electro- and chemically competent E. coli

CopyCutter™ EPI400™ E.coli* cells were developed to significantly lower the copy number of a wide variety of common vectors so that you can more readily clone unstable DNA sequences

BioSearch Tech (Lucigen/Epicentre)

Catalogue No.DescriptionPack SizePriceQty
C400CH10CopyCutter™ EPI400™ Chemically Competent E.coli 10 x 50 µl £173.00£173.00Offer until : 31-Dec-2020Contact Cambio for special pricing on all orders over £2000 View Offer Quantity Add to Order
C400EL10CopyCutter™ EPI400™ Electrocompetent E.coli 10 x 50 µl £224.00£224.00Offer until : 31-Dec-2020Contact Cambio for special pricing on all orders over £2000 View Offer Quantity Add to Order
CIS40025CopyCutter™ Induction Solution25 ml £85.00£85.00Offer until : 31-Dec-2020Contact Cambio for special pricing on all orders over £2000 View Offer Quantity Add to Order

CopyCutter™ EPI400™ Electro- and chemically competent E. coli

CopyCutter™ EPI400™ E.coli* cells were developed to significantly lower the copy number of a wide variety of common vectors so that you can more readily clone unstable DNA sequences

BioSearch Tech (Lucigen/Epicentre)

CopyCutter™ EPI400™ E.coli* cells were developed to significantly lower the copy number of a wide variety of common vectors so that you can more readily clone unstable DNA sequences. 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 (Figure 1).

The CopyCutter™ EPI400™ cell line was derived from our high-transformation efficiency phage T1-resistant TransforMax™ EC100™ 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. Moreover, following a short incubation in the presence of the CopyCutter™ Induction Solution, you can increase the copy number of the vector to improve plasmid yields (Figure 2).

*Covered by issued and/or pending patents.

Figure 1. DNA inserts encoding toxic gene products were successfully cloned into high-copy number vectors using CopyCutter EPI400 E. coli cells Figure 1. DNA inserts encoding toxic gene products were successfully cloned into high-copy number vectors using CopyCutter EPI400 E. coli cells
Figure 1. DNA inserts encoding toxic gene products were successfully cloned into high-copy number vectors using CopyCutter™ EPI400™ E. coli cells After sequencing, the full-length acpP clones in TransforMAX™ EC100™ cells were found to contain multiple point mutations. Figure 2. Uninduced CopyCutter™ EPI400™ E. coli cells containing a regB clone (Lane U) are induced to higher-copy number (Lane I) using the CopyCutter™ Induction Solution. Crude extracts of plasmid DNA were prepared from cells grown in selective media and analyzed by agarose gel electrophoresis. Approximately the same number of lysed cells (based on OD600) were loaded per lane.

 

Genotype

F- mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL nupG trfA tonA pcnB4 dhfr

CopyCutter™ EPI400™ Electrocompetent E. coli

Transformation efficiency of >1 X 1010 cfu/µg of pUC19.

  • Supplied in convenient single-use 50-µl volumes.

 

CopyCutter™ EPI400™ Chemically Competent E. coli

Transformation efficiency of >1 x 108 cfu/µg of pUC19.

  • Supplied in convenient single-use 50-µl volumes.

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

CopyCutter™ EPI400™ Electro- and chemically competent E. coli

CopyCutter™ EPI400™ E.coli* cells were developed to significantly lower the copy number of a wide variety of common vectors so that you can more readily clone unstable DNA sequences

BioSearch Tech (Lucigen/Epicentre)

Protocols for: CopyCutter™ EPI400™ Electrocompetent E. coli

Due to the constant updating of the protocols by the manufacturer we have provided a direct link to Lucigen’s product page, where the latest protocol is available.

Please note this will open a new page or window on your computer.

 CopyCutter™ Protocol

(catalogue number C400EL10 / C400CH10 / CIS40025)

Please note: all protocols off site are the responsibility of the products supplier

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

CopyCutter™ EPI400™ Electro- and chemically competent E. coli

CopyCutter™ EPI400™ E.coli* cells were developed to significantly lower the copy number of a wide variety of common vectors so that you can more readily clone unstable DNA sequences

BioSearch Tech (Lucigen/Epicentre)

References

 

  1. Bryan, J. D. & Shelver, D. W. (2008) Streptococcus agalactiae CspA Is a Serine Protease That Inactivates Chemokines, Science 320 , 1230-.
  2. Grefen, C., et al. (2008) Subcellular Localization and In Vivo Interactions of the Arabidopsis thaliana Ethylene Receptor Family Members, Mol Plant 1 , 308-320.
  3. Schroeder, L. K., et al. (2007) Function of the Caenorhabditis elegans ABC Transporter PGP-2 in the Biogenesis of a Lysosome-related Fat Storage Organelle, Mol. Biol. Cell 18 , 995-1008.

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

CopyCutter™ EPI400™ Electro- and chemically competent E. coli

CopyCutter™ EPI400™ E.coli* cells were developed to significantly lower the copy number of a wide variety of common vectors so that you can more readily clone unstable DNA sequences

BioSearch Tech (Lucigen/Epicentre)

Applications

  • CRISPR and cloning of large or difficult fragments

  • Cloning of unstable DNA sequences or those expressing toxic proteins.

Benefits

  • Maintain clones at low-copy number, then induce to higher copy number for improved plasmid yield.
  • High transformation efficiency with clones of all sizes.
  • Resistance to bacteriophages T1 and T5.
  • Enables efficient cloning of methylated DNA.
  • High yields of DNA.
  • Stability of large cloned inserts.

 

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200