Bacterial transformation is a common technique in molecular biology with a variety of applications, such as DNA sequencing, library construction or cloning. Transformation requires the use of competent bacterial cells, which have been treated to allow plasmid DNA to enter into the cells’ cytoplasm. They are commonly either electrocompetent or chemically competent.
Wel-bioAps’ chemically competent cells have been treated with calcium chloride to neutralise the negative charges of both the phospholipid bilayer of the cell membrane and the plasmid DNA, which allows the DNA to approach the cell membrane. The cells are then heat-shocked to render the membrane more permeable, which allows the plasmid DNA to travel into the cytoplasm. All competent cells and been thoroughly assessed and have industry-leading transfection rates.
BL21(DE3)pLysS
- Predominantly used for protein expression.
- Wel-bioAps BL21(DE3)pLysS competent cells are deficient in both lon protease, and the OmpT outer membrane protease, which both decrease heterologous protein expression degradation.
- They carry the λDE3 prophage which contains the T7 polymerase gene under a lacUV5 promoter, so IPTG can be used to express T7 RNA polymerase.
- In pLysS strains, pET recombinants which encode target proteins are stabilised by suppressing unwanted basal T7 RNA polymerase activity with T7 lysosyme expression.
- Transfection efficiency - TBD cfu/ml.
DH5a(DE3)
- Predominantly used for plasmid transformation protocols.
- DH5a cell strains contain recA1, endA1 and lacZΔM15 mutations which stabilise and enhance transformations, and enable blue-white screening of transformed cells.
- They carry the λDE3 prophage which contains the T7 polymerase gene under a lacUV5 promoter, so IPTG can be used to express T7 RNA polymerase.
- Transfection efficiency - TBD cfu/ml.
How to order
- We work closely with our customers to provide competent cells that meet the most exacting requirements.
- To begin the process, please email orders@wel-bioaps.com with any special requirements you may have.
- One of our team of scientists will contact you as soon as possible to discuss your needs and provide details on the next steps.
