RED/ET RECOMBINATION KITS GENE BRIDGES
E.coli Strain Modification

With Red/ET you can easily modify the E. coli genome:

• gene disruption, deletion or insertion
• reporter gene and tag integration
• promoter fine tuning
• introduction of point mutations

Red/ET Recombination Kits

PRed/ET allows for the genetic engineering of tailor-made targeting constructs for animal models:

• conditional knock-out/knock-in
• promoter or reporter fusions
• exon swapping
• introduction of point mutations

Red/ET Recombination Kits

SELECTION CASSETTES GENE BRIDGES
Additional selection marker cassettes

PLASMIDS and STRAINS GENE BRIDGES
Modification Experiments

Increase your flexibility by using additional selection marker cassettes optimized for Red/ET recombination.

• Selection cassettes are driven by eukaryotic (PGK) and prokaryotic (gb2) promoters, respectively
• Selection markers flanked by loxP or FRT-sites can be removed in a Cre or Flp recombination step where appropriate
• Due to a modular architecture, selection cassettes can be PCRamplified with master primers
• Zero background because cassettes are encoded by suicide plasmids to avoid false positive clones

Selection Cassettes

The plasmids and strains are very useful for verifying the functionality of loxP- or FRT-sites within a DNA construct or for excising FRT- or loxP-flanked DNA stretches (such as selection markers) during a modification task:

* The plasmids are easily transformed into cells resulting in an activation of the recombinases in vivo.
* The E.coli strains can be used as deleter strains, into which DNA can be transformed.

Plasmids and Strains

Red/ET Recombination from Gene Bridges allows a faster, more flexible and highly reliable modification of plasmids, BACs, or the E. coli genome than conventional cloning methods. Red/ET exploits phage l homologous recombination potential for in vivo genetic engineering in E. coli.

Since Red/ET does not depend on restriction enzymes, ligation reactions or in vitro cleanup steps, it is highly applicable for the engineering of large DNA molecules.

Gene Bridges Red/ET Recombination is a revolutionary method for DNA engineering. Recombineering with Red/ET allows unlimited cloning, subcloning, and modification of DNA at any chosen position. It permits precise engineering of DNA molecules of any size, including very large ones such as BACs or the E.coli chromosome.
Advantages over conventional methods
* Independent of restriction sites
* No size limits
* No unwanted mutations
* Rapid
Red/ET Recombineering is one simple and powerful tool to modify plasmids, BACs, and even E.coli chromosomes.

• only 50 bp of flanking sequence sufficient for recombination
• sequence independent
• precise at any position
• cloning without restriction enzymes
• no ligation reactions
• cloning of inserts up to 80 kb

Cells which express l-derived red genes from plasmid pRed/ET promote base precise exchange of DNA sequences flanked by homology arms. The in vivo reaction is catalyzed by the exonuclease Reda and the DNA annealing protein Redb. Even the most demanding tasks can be reduced to three basic steps:
   1. Attachment of Homology Arms
   2. Recombineering
   3. Selection/Screening