Bacterial Transformation Service
Bacterial transformation is an important method in molecular biology and genetics research. It is a process of horizontal gene transfer (the transfer of exogenous genetic material from one bacterium to another) that occurs in bacterial cells. Through the transformation process, exogenous genetic material can be introduced into bacterial cells, and through the replication and expression of exogenous DNA, new genetic traits can appear in recipient cells.
Bacterial Transformation
In prokaryotes, transformation is a very common life phenomenon. The transformed bacteria can acquire new genetic characteristics and phenotypes. In 1928, the transformation was first discovered by Frederick Griffith in Streptococcus pneumoniae, and in 1944, Oswald Avery, Colin MacLeod and Maclyn McCarty proved the principle of transformation. When using a plasmid vector to transform bacteria, it is necessary to induce the recipient bacteria to produce a short-term competence to take in foreign DNA.
Figure 1. Bacterial transformation techniques (Ren.; et al. 2019)
Competent Cell
Competent state refers to a specific physiological state when the recipient cell is susceptible to foreign DNA. Competence is affected by the external environment. In the laboratory, bacteria (such as Escherichia coli) are generally processed by physical and chemical methods to obtain competent cells. CaCl2 solution treatment is a common chemical method, which has the advantage of simple operation and the conversion efficiency can also meet the requirements of general experiments. Electroporation is a commonly used physical method for preparing competent cells. In a competent cell, the permeability of the cell membrane is greatly increased, making it easy for foreign DNA molecules to enter the cell from the gaps on the membrane surface.
We also provide customers with many types of competent cell products. If you want to purchase competent cells, please feel free to contact us for product specific information.
Bacterial Transformation Process
- Preparation of competent cells
A single colony of E. coli is selected and cultured in a liquid medium to the logarithmic growth phase, and then processed by heat-shock transformation or electroporation to obtain competent cells. Competent cells can be aliquoted and stored at -70°C for a long time.
- Transformation
- Mix competent cells and plasmids (mix at the ratio of 100μL competent cells and 10ng plasmids). After standing on ice for 20-30 minutes, in a water bath at 42°C for 30-60 seconds, then quickly put it back on ice and let it stand for 2-5 minutes.
- Add 500μL of antibiotic-free medium, mix well, and incubate for 1h in a shaker at 37°C to activate and express the resistance gene.
- Take the cultured bacteria solution and inoculate it on a plate containing the corresponding antibiotics, and incubate overnight at 37°C.
- After overnight incubation, colonies appear in the plate, observe and record. Only bacteria containing resistance genes can grow on antibiotic-containing plates.
Workflow of Bacterial Transformation Service at Creative BioMart Microbe
Creative BioMart Microbe provides bacterial transformation services with high transformation efficiency. The materials involved in transformation experiments can be provided by us or obtained through our services. You can also choose to provide your own materials and have our technicians perform bacterial transformation experiments.
If you want to know more about this service, please feel free to contact us by phone or email, or you can consult through our inquiry form. We will ensure to provide you with efficient and high-quality experimental outsourcing services.
Reference
- Ren, J.; et al. Artificial transformation methodologies for improving the efficiency of plasmid DNA transformation and simplifying its use. Applied Microbiology and Biotechnology. 2019, 103: 9205-9215.
