Stability
The competent cells should be viable for three to six months from the date of manufacture when stored as instructed.
Activity
The Kit is tested to see competent cells of E.coli strain transformed as instructed in the protocol
Description
Transformation, a process central to recombinant DNA technology, is the process by which a foreign DNA is introduced into a bacterium. In this kit, the experiment employs a 3-5kb circular Plasmid DNA as the foreign DNA. The Plasmid contains genes of interest to us along with a selection markers (i.e., genes which confer antibiotic resistance). For Transformation to be successful, the recipient bacterium, called the Host, should first be made Competent for the uptake of DNA. This kit describes a simple procedure for the same. The Competent status, however, is not permanent and is lost eventually. In general, freshly made Competent cells provide high Transformation efficiency. The molecular mechanism for blue-white screening is based on the Lac Operon. The chemical required for this screening is X-Gal. It is the hydrolysis of X-Gal that causes the characteristic blue color.
Includes
| Components | 15 Experiments | 30 Experiments |
|---|
| Host cells | 1.0 ml | 2.0 ml |
| Sterile LB medium | 50.0 ml | 60.0 ml |
| Sterile LB medium | 10.0 ml | 15.0 ml |
| Competent cells | 3.0 ml | 5.0 ml |
| Plasmid DNA (50 ng/mcl) | 40.0 mcl | 80.0 mcl |
| 0.1M CaCl2 | 30.0 ml | 60.0 ml |
| LB agar powde | 18.0 g | 36.0 g |
| Ampicillin | 400 mg | 800 mg |
| X-Gal (20mg/ml) | 1.0 ml | 2.0 ml |
| IPTG (200 mg/ml) | 150 mcl | 300 mcl |
| Sterile water | 1.0 ml | 2.0 ml |
