Red/ET Quick & Easy BAC Modification Kit (need license to buy)
This kit is designed to modify BACs with the revolutionary Red/ET Recombination technology - a powerful tool for introducing modifications such as insertions and deletions into any type of BAC within 2 weeks. Red/ET Recombination uses in vivo recombination and proofreading activity to minimize unwanted mutations in your construct. Included in the kit are all the necessary contents for modifying BACs and all materials needed to perform a control experiment intended to familiarize you with this novel technique. Please Note: This product is only available to licensed customers. Academic and non-profit laboratories can obtain a license free-of-charge by simply signing off on the terms of the Red-ET Non-Commercial License (.pdf file can be found on the right column of this page). Companies need to contact Gene Bridges directly regarding licensing terms (link can be found under Suppliers). Please contact B-Bridge International or Gene Bridges for further information.
- The E. coli strain carrying the BAC, which is to be modified, is transformed with the expression plasmid pRedET.
- The expression of genes mediating Red/ET is induced by the addition of Larabinose and a temperature shift from 30C to 37C . After induction the cells are prepared for electroporation and the linear Tn5-neo cassette (PCR product) flanked by homology arms is electroporated. Red/ET recombination inserts the functional cassette into the target locus. Only colonies carrying the modified BAC will survive Kanamycin selection on the agar plates.
- The successful integration of the selection cassette will be monitored by PCR or DNA mini preparation.