Conditional Knockout

Probably the easiest and most convenient method of generating targeting vectors is to let Gene Bridges do it for you. We apply Red/ET Recombineering, a novel DNA engineering technology, to generate targeting vectors which fit your requirements precisely - nucleotide precisely.

Conditional Knock-out Constructs for € 9,990
To enquire about ordering conditional knock-outs constructs, please download the Conditional Knockout Form.

For information on all DNA engineering services offered by Gene Bridges see General DNA Engineering.

Conditional Knock-out constructs made by Red/ET Recombineering include:

• Identifying and ordering two appropriate BACs
• Sub-cloning of a fragment up to 18kb from a BAC into a high copy vector system provided by Gene Bridges
• Insertion of a single loxP site into one location of the sub-cloned DNA
• Insertion of a FRT-PGK-gb2-neo-FRT-loxP cassette (kanamycin/neomycin resistance cassette under the control of a prokaryotic and a eukaryotic promoter) into a second location of the sub-cloned DNA yielding the final targeting vector
• Verification check of all FRT, loxP sites and PGK promoter by sequencing
• Estimated timelines: 12 weeks from receipt of ordered BACs

You provide:

• Exact information about the gene to be targeted incl. accession number
• Length of desired homology arms to be inserted upstream and downstream of the functional elements to be inserted (e.g. 12kb upstream and 5kb downstream). Please indicate the maximum length of the locus to be sub-cloned (limited to 18kb)

You can choose:

• 1-2 additional restriction sites flanking the locus to be sub-cloned for later linearization of the targeting plasmid
• Exact insertion position of single loxP site (between nt A and nt B)
• Exact insertion position of the FRT-PGK-gb2-neo-FRT-loxP (between nt C and nt D)