TA0100
Sigma
TargeTron® Gene Knockout System
Description
| Frequently Asked Questions | Live Chat and Frequently Asked Questions are available for this Product. |
| Components | System User Manual TargeTron Access card (password for on-line target site selection) Intron PCR Template EBS Universal Primer LacZ Control Primers pACD4K-C Linear Vector (20 ng/μl) JumpStart™ REDTaq® ReadyMix® HindIII 20 U/μl BsrGI 10 U/μl 10× Restriction Enzyme Buffer |
| Features and Benefits | Targeted and permanent gene disruption Simple, streamlined protocol; Knockouts in 3 days or less Minimal screening to isolate mutants >90% successful targeted insertion No cell conjugation or specific host factor requirements |
| General description | For detailed information, please visit the TargeTron® home page: www.sigma-aldrich.com/targetron. The TargeTron 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 as Escherichia coli, Staphylococcus aureus, Clostridium perfringens, Shigella flexneri, Salmonella typhimurium, and Lactococcus 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 (3 EA or 10 EA). 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 3 EA kit are sufficient for 3 designs, 12 reactions. Reagents in the 10 EA kit are sufficient for 10 designs, 40 reactions. |
| Other Notes | To view the TargeTron Gene Knockout System animation, visit www.sigma-aldrich.com/targetronanimation. |
| Legal Information | This product and its use are the subject of one or more of U.S. Patent Nos. 5,698,421, 5,804,418, 5,869,634, 6,027,895, 6,001,608, and 6,306,596 and/or other pending U.S. and foreign patent applications controlled by InGex, LLC. |
| REDTaq is a registered trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co. | |
| JumpStart is a trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co. | |
| TargeTron is a registered trademark of InGex, LLC |
Properties
| shipped in | wet ice |
| storage temp. | −20°C |
Safety
| Risk Statements | 25-36/37/38 |
| Safety Statements | 26-36-45 |
Related Products
| Related product | G5916, Gene Knockout Protocols |
