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.
Origami™ 2 host strains are K-12 derivatives that have mutations in both the thioredoxin reductase (trxB) and glutathione reductase (gor) genes, which greatly enhance disulfide bond formation in the E. coli cytoplasm. The Origami™ 2 strains are kanamycin sensitive; like the original Origami™ strains, the gor mutation is still selected for by tetracycline. To reduce the possibility of disulfide bond formation between molecules, strains containing mutations in trxB and gor are recommended only for the expression of proteins that require disulfide bond formation for proper folding.
