|Products||TargeTron Gene Knockout System from Sigma-Aldrich|
|Item||TargeTron Gene Knockout System|
|Description|| The TargeTron(TM) Gene Knockout System provides optimized reagents and protocols for the rapid and specific disruption of bacterial genes by insertion of group II introns. The method exploits the retrohoming ability of group II introns and utilizes a simple PCR step to re-target the TargeTron group II intron for specific insertion into the host genome. Utility of the technology has been demonstrated for prokaryotic genetic engineering, systems biology and functional genomics approaches.|
Compared to existing knockout methodologies, the TargeTron system is quite rapid, simple and robust. The process involves loading the target gene sequence into the well-validated TargeTron algorithm on the Sigma website (www.sigma-aldrich.com/targetronaccess), using one or more of the output primer sets to mutate the TargeTron intron template via PCR, ligation of the fragment into the pACD4K-C linear vector, transformation of the host, induction of gene disruption, selection by kanamycin resistance (insertion of activated marker) and screening for the desired knockout.
Gene knockout using the TargeTron system has been validated in a broad range of bacterial strains such asEscherichia coli, Staphylococcus aureus, Clostridium perfringens, Shigella flexneri, Salmonella typhimurium,andLactococcus lactis. The system may be modified for use in additional organisms.
Each TargeTron kit provides an access card and password for a specified number of designs (3EA or 10EA). The password gives the user access to the extensively validated TargeTron algorithm that predicts optimal intron insertion sites and designs primers for mutating the intron to insert into those sites. After selection of one or more desirable insertion points, the required oligos are ordered to begin laboratory procedures using the kit. Reagents in the 3EA kit are sufficient for 3 designs, 12 reactions. Reagents in the 10EA kit are sufficient for 10 designs, 40 reactions.
St. Louis, MO, USA
Customer Service: 800-325-3010
Tech Support: 800-325-5832
Fax Number: 314-771-5757
Web Site: http://www.sigmaaldrich.com/