A common method for transformation of DNA plasmids into E. coli is the use of chemically competent cells. Although competent cells can be prepared in the laboratory, greater efficiency, reproducibility, and convenience are achieved using Novagen prepared competent cells. Novagen® Singles Competent Cells are designed for ultimate convenience and reliability in plasmid transformation. Chemically competent cells are prepared using an optimized method. Cells are provided in 50µl volumes, eliminating the need to aliquot, freeze/thaw, or waste partially used vials, saving time and money and ensuring reliable cell performance. To use, simply thaw, add DNA, incubate 5 minutes on ice, heat shock for 30 seconds, place on ice for 2 minutes, and add SOC medium.
T7 expression strains are lysogens of bacteriophageDE3, as indicated by the (DE3). These hosts carry achromosomal copy of the T7 RNA polymerase gene under control of the lacUV5 promoter. Such strains are suitable for production of protein from target genes cloned in appropriate T7 expression vectors, using IPTG as an inducer.
Rosetta 2 host strains are BL21 derivatives designed to enhance the expression of eukaryotic proteins that contain codons rarely used in E. coli. These strains supply tRNAs for 7 rare codones (AGA, AGG, AUA, CUA, GGA, CCC, and CGG) on a compatible chloramphenicol-resistant plasmid. The tRNA genes are driven by their native promoters.
DE3 indicates that the host is a lysogen of λDE3, and therefore carries a chromosomal copy of the T7 RNA polymerase gene under control of the lacUV5 promoter. Such strains are suitable for production of protein from target genes cloned in pET vectors by induction with IPTG.
pLysS strains express T7 lysozyme, which further suppresses basal expression of T7 RNA polymerase prior to induction, thus stabilizing pET recombinants encoding target proteins that affect cell growth and viability.
Novagen′s Rosetta 2 (pLysS) host strains are BL21 derivatives designed to enhance the expression of eukaryotic proteins that contain codons rarely used in E. coli along with T7 lysozyme activity.
Genotype: F-ompT hsdSB(rB- mB-) gal dcm (DE3) pLysSRARE2 (CamR)
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Features and Benefits
• Guaranteed transformation efficiencies:
>1.5 × 10⁸ cfu/µg (NovaBlue andNovaBlueT1R)
>2 × 10⁶ cfu/µg [BL21(DE3), BL21(DE3)pLysS,Origami(DE3), Origami 2(DE3),Rosetta(DE3), Rosetta 2(DE3)pLysS]
• Provided as frozen single-use aliquots in 11-reaction or 22-reaction kits
• Includes Test Plasmid and SOC Medium
• Selection of popular host strains: NovaBlue forhigh efficiency cloning, and λDE3 lysogens forprotein expression with pET vectors
• Prepared using an optimized chemical method