TransforMax™ EPI300™ E. coli cells lack the tonA gene and are engineered for use with EPICENTRE's CopyControl™ cDNA, Gene, and PCR Cloning Kit and other CopyControl Cloning Systems* that do not require phage T1-resistant cells. The cells contain an inducible mutant trfA gene whose gene product is required for initiation of replication from the oriV origin of replication, such as that in CopyControl pCC1™ vectors or in clones that are retrofitted with CopyControl capability using the EZ-Tn5™<oriV/KAN-2> Insertion Kit. On LB chloramphenicol plates or in LB or SOC media supplemented with chloramphenicol, CopyControl clones grown in TransforMax EPI300 E. coli replicate at single-copy number from the F-factor replicon because expression of the trfA gene is repressed. Addition of CopyControl Induction Solution induces the cells to express the trfA gene product and induces their replication at high-copy number from oriV (Fig. 1). Replication from oriV results in higher yields and higher purity of cloned DNA.

Genotype

F^- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ^- rpsL (Str^R) nupG trfA dhfr

TransforMax EPI300 Electrocompetent E. coli

• Transformation efficiency of >1 x 10¹⁰ cfu/µg of pUC19.

TransforMax EPI300 Chemically Competent E. coli

• Transformation efficiency of >5 x 10⁸ cfu/µg of pUC19.

For use with vectors or clones having an inducible oriV for CopyControl™ capability.

 *Covered by issued and/or pending patents.

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Protocol

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Citations

TransforMax™ EPI300™ Electrocompetent E. coli
TransforMax™ EPI300™ Chemically Competent E. coli

1. Amore, G., et al. (2010) Multi-year evolutionary dynamics of West Nile virus in suburban Chicago, USA, 2005-2007, Phil Trans R Soc B 365 , 1871-1878.
2. Benders, G. A., et al. (2010) Cloning whole bacterial genomes in yeast, Nucleic Acids Res. 38 , 2558-2569.
3. Jin, Y. O., et al. (2010) Association of Missense Mutations in Epoxyalkane Coenzyme M Transferase with Adaptation of Mycobacterium sp. Strain JS623 to Growth on Vinyl Chloride, Appl. Envir. Microbiol. 76 , 3413-3419.
4. Oh, J., et al. (2010) A universal TagModule collection for parallel genetic analysis of microorganisms, Nucleic Acids Res.  , gkq419.
5. Witte, B., et al. (2010) Functional Prokaryotic RubisCO from an Oceanic Metagenomic Library, Appl. Envir. Microbiol.  , AEM.02661-09.
6. Allender, C. J., et al. (2009) Identifying the Source of Unknown Microcystin Genes and Predicting Microcystin Variants by Comparing Genes within Uncultured Cyanobacterial Cells, Appl. Envir. Microbiol. 75 , 3598-3604.
7. Balder, R., et al. (2009) Hag Mediates Adherence of Moraxella catarrhalis to Ciliated Human Airway Cells, Infect. Immun. 77 , 4597-4608.
8. Gibson, D. G. (2009) Synthesis of DNA fragments in yeast by one-step assembly of overlapping oligonucleotides, Nucleic Acids Res. 37 , 6984-6990.
9. Yung, P. Y., et al. (2009) Phylogenetic screening of a bacterial, metagenomic library using homing endonuclease restriction and marker insertion, Nucleic Acids Res. 37 , e144-.
10. Zhang, X., et al. (2009) A One-Plasmid System To Generate Influenza Virus in Cultured Chicken Cells for Potential Use in Influenza Vaccine, J. Virol. 83 , 9296-9303.
11. Sen, K., et al. (2007) Development of an Internal Control for Evaluation and Standardization of a Quantitative PCR Assay for Detection of Helicobacter pylori in Drinking Water, Appl. Envir. Microbiol. 73 , 7380-7387.
12. Roberts, A. P., et al. (2006) Characterization of the Ends and Target Site of a Novel Tetracycline Resistance-Encoding Conjugative Transposon from Enterococcus faecium 664.1H1, J. Bacteriol. 188 , 4356-4361.
13. Hearnes, J. M., et al. (2005) Chromatin Immunoprecipitation-Based Screen To Identify Functional Genomic Binding Sites for Sequence-Specific Transactivators, Mol. Cell. Biol. 25 , 10148-10158.
14. Ratnayaka, I., et al. (2005) Construction and Characterization of a BAC Library of a Cold-Tolerant Hexaploid Wheat Cultivar, Crop Sci. 45 , 1571-1577.
15. Han, S. O., et al. (2004) Isolation and Expression of the xynB Gene and Its Product, XynB, a Consistent Component of the Clostridium cellulovorans Cellulosome, J. Bacteriol. 186 , 8347-8355.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Applications

• CRISPR and cloning of large or difficult fragments
• Generation of inducible copy-number clones using the CopyControl™ Cloning System.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200