{"product_id":"copycutter-epi400-competent-cells-comcel-010","title":"CopyCutter EPI400 Competent Cells","description":"\u003cp\u003eCopyCutter™ 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. 2).1\u003c\/p\u003e\u003cp\u003eApplications\u003c\/p\u003e\u003cp\u003eCloning of unstable DNA sequences or those expressing toxic proteins.\u003c\/p\u003e\u003cp\u003eThe CopyCutter EPI400 cell line was derived from Epicentre's high-transformation efficiency phage T1-resistant TransforMAX™ EC100™-T1RE.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.\u003c\/p\u003e\u003cp\u003eThe 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, a short incubation in the presence of the CopyCutter Induction Solution can increase the copy number of the vector to improve plasmid yields (Figure. 3).\u003c\/p\u003e\u003cp\u003eFigure 1. Copy-number of ColE1-type plasmids is lowered up to 25-fold in CopyCutter EPI400 E. coli cells. Lanes C, TransforMax™ EC100™ cells; Lanes U and I, uninduced or induced CopyCutter EPI400 cells. DNA extracts from the same number of lysed cells (based on OD600) were loaded per lane.\u003c\/p\u003e\u003cp\u003eFigure 2. 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.\tFigure 3. 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 analysed by agarose gel electrophoresis. Approximately the same number of lysed cells (based on A600) were loaded per lane.\u003c\/p\u003e\u003cp\u003eBenefits\u003c\/p\u003e\u003cp\u003eMaintain clones at low-copy number, then induce to higher copy number for improved plasmid yield.\u003cbr\u003eHigh transformation efficiency with clones of all sizes.\u003cbr\u003etonA for resistance to bacteriophages T1 and T5.\u003cbr\u003elacZΔM15 for blue\/white screening of recombinants.\u003cbr\u003eRestriction minus [mcrA, Δ(mrr-hsdRMS-mcrBC)] genotype enables efficient cloning of methylated DNA.\u003cbr\u003eEndonuclease minus (endA1) to ensure high yields of DNA.\u003cbr\u003eRecombination minus (recA1) for greater stability of large cloned inserts.\u003c\/p\u003e\u003cp\u003eGenotype\u003c\/p\u003e\u003cp\u003eF- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL (StrR) nupG trfA tonA pcnB4 dhfr\u003c\/p\u003e\u003cp\u003eCopyCutter EPI400 Electrocompetent E. coli\u003c\/p\u003e\u003cp\u003eTransformation efficiency of \u0026gt;1 x 1010 cfu\/µg of pUC19.\u003c\/p\u003e\u003cp\u003eCopyCutter EPI400 Chemically Competent E. coli\u003c\/p\u003e\u003cp\u003eTransformation efficiency of \u0026gt;1 x 107 cfu\/µg of pUC19.\u003c\/p\u003e\u003cp\u003eReference\u003c\/p\u003e\u003cp\u003eHaskins, D. (2004) Epicentre Forum 11(5), 6.\u003c\/p\u003e\u003cp\u003e*Covered by issued and\/or pending patents.\u003c\/p\u003e\u003cp\u003eread more\u003c\/p\u003e\n\u003ch2\u003eKey features\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003eOptimised for cloning toxic or unstable DNA\u003c\/li\u003e\n\u003cli\u003eStabilise toxic inserts in common cloning and expression vectors (pUC and pET-type vectors)\u003c\/li\u003e\n\u003cli\u003eClone and maintain challenging sequences at reduced plasmid copy number, then induce to high copy number for DNA recovery\u003c\/li\u003e\n\u003cli\u003eAvoid T1 and T5 phage contamination with tonA mutation\u003c\/li\u003e\n\u003cli\u003eChoose chemically competent cells for general cloning or electrocompetent cells for demanding applications such as library generation\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003ch2\u003eProduct Details\u003c\/h2\u003e\n\u003cp\u003eTen tubes each containing 50 µL of cells (enough cells for 10 transformations), CopyCutter Induction Solution and pUC19 control DNA. The CopyCutterInduction Solution is provided at a 1,000X concentration and is filter sterilised.\u003c\/p\u003e\n\u003ch2\u003eDocuments\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/sds_COMCEL-010_C400CH10-CopyCutter-EPI400-Chemically-Competent-E-coli\"\u003eC400CH10 CopyCutter EPI400 Chemically Competent E coli\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/sds_COMCEL-010_C400EL10-CopyCutter-EPI400-Electrocompetent-Ecoli\"\u003eC400EL10 CopyCutter EPI400 Electrocompetent Ecoli\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/manual_COMCEL-010_CopyCutter-cells-\u0026amp;-Induction-Soln\"\u003eCopyCutter cells \u0026amp; Induction Soln\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/pis_COMCEL-010_F306-US-CopyCutter-EPIT400-Comp-Cells-X-1a\"\u003eF306 US CopyCutter EPIT400 Comp Cells X 1a\u003c\/a\u003e\u003c\/li\u003e\n\u003c\/ul\u003e","brand":"Biosearch Technologies","offers":[{"title":"10 Reactions","offer_id":45986439397455,"sku":"C400CH10","price":1250.0,"currency_code":"ILS","in_stock":true},{"title":"10 Reactions (C400EL10)","offer_id":45986439430223,"sku":"C400EL10","price":1658.0,"currency_code":"ILS","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0712\/0748\/5519\/files\/CopyCutter-EPI400-Competent-Cells_df34fd19-d77c-45f5-8221-749046b92c12.jpg?v=1785956183","url":"https:\/\/www.danyel.co.il\/products\/copycutter-epi400-competent-cells-comcel-010","provider":"danyel biotech","version":"1.0","type":"link"}