A common method for transformation of 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. Every Novagen® competent cell strain is verified for phenotype and purity, and is guaranteed for transformation efficiency.
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 stabilising pET recombinants encoding target proteins that affect cell growth and viability.
The standard volume per tube is 0,2 ml, which provides enough cells for 10 transformations. All kits are supplied with SOC medium and test plasmid. Several strains are also available as Singles™ Competent Cells, which are provided in single-use, 50 μl volumes for extra convenience and efficiency.
Tuner™ strains are lacZY deletion mutants of BL21 and enable adjustable levels of protein expression throughout all cells in a culture. The lac permease (lacY) mutation allows uniform entry of IPTG into all cells in the population, which produces a concentration-dependent, homogeneous level of induction. By adjusting the concentration of IPTG, expression can be regulated from very low levels up to the robust, fully induced levels commonly associated with pET vectors. Lower-level expression may enhance the solubility and activity of difficult target proteins.