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

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Applications

  • Generation of inducible copy-number clones using the CopyControl™ Cloning System.

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.

Benefits

  • Copy-number of clones is under tight control of an inducible promoter linked to the trfA gene.
  • High transformation efficiency with clones of all sizes.
  • lacZΔM15 for blue/white screening of recombinants.
  • Restriction-minus [mcrA, Δ(mrr-hsdRMS-mcrBC)] phenotype enables efficient cloning of methylated DNA.
  • Endonuclease-minus (endA1), to ensure high yields of DNA.
  • Recombination-minus (recA1), for greater stability of large insert clones.

Figure 1

Figure 1. Copy number of CopyControl™ BAC clones can be induced 10- to 20-fold in TransforMax™ EPI300™ E. coli. The yield of BAC DNA from CopyControl BAC clones of 110-145 kb increased >14-fold following addition of CopyControl Induction Solution. U=uninduced cells; I=induced cells.

Genotype

F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ- rpsL (StrR) 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.

*Covered by issued and/or pending patents.


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   Catalog No. Concentration Size

TransforMax™ EPI300™ Electrocompetent E. coli
   EC300105 5 X 100 µl
   EC300110 10 X 100 µl
   EC300150 50 X 100 µl
Includes CopyControl™ Induction Solution and pUC19 control DNA.

TransforMax™ EPI300™ Chemically Competent E. coli
   C300C105 10 X 50 µl
Includes CopyControl™ Induction Solution and pUC19 control DNA.

CopyControl™ Induction Solution
   CCIS125 25 ml
1,000X concentrated solution. Filter sterilized.

You may wish to consider the following related products:

Colony Fast-Screen™ Kit (PCR Screen)
Colony Fast-Screen™ Kit (Restriction Screen)
Colony Fast-Screen™ Kit (Size Screen)
CopyControl™ BAC Cloning Kits (BamH I, EcoR I, or Hind III)
CopyControl™ pCC1BAC™ Cloning-Ready Vectors (BamH I, EcoR I, or Hind III)

CopyControl™ cDNA, Gene and PCR Cloning Kits
CopyControl™ Fosmid Library Production Kit
CopyControl™ HTP Fosmid Library Production Kit

Electroporation Cuvettes
EZ-Tn5™ <oriV/KAN-2> Insertion Kit
TransforMax™ EPI300™-T1R Electrocompetent E. coli
TransforMax™ EPI300™-T1R Chemically Competent E. coli

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