Components

11 rxn or 22 rxnRosetta 2(DE3)pLysS Competent Cellsr> 2 × 2 ml or 4 × 2 mlSOC Mediumr> 10 µlTest Plasmid

Features and Benefits

Guaranteed transformation efficiencies:r>>1.5 × 10⁸ cfu/µg (NovaBlue andNovaBlueT1R)r>>2 × 10⁶ cfu/µg [BL21(DE3), BL21(DE3)pLysS,Origami™(DE3), Origami 2(DE3),Rosetta™(DE3), Rosetta 2(DE3)pLysS]r> Provided as frozen single-use aliquots in 11-reaction or 22-reaction kitsr> Includes Test Plasmid and SOC Mediumr> Selection of popular host strains: NovaBlue forhigh efficiency cloning, and λDE3 lysogens forprotein expression with pET vectorsr> Prepared using an optimized chemical method

General description

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 hsdS_B(r_B^- m_B^-) gal dcm (DE3) pLysSRARE2 (Cam^R)r>r>r>This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges us to request certain information about you and the establishment where the GMOs are being handled. Click here for Enduser Declaration (EUD) Form.

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.

Legal Information

NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany

Packaging

22 reactions in Glass bottle

11 reactions in Plastic ampoule

Warning

Toxicity: Multiple Toxicity Values, refer to MSDS (O)