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requests that are tailored to your specific need. Send any question to KOserv@cellogenetics.com To request this service, please download
the order form here.
BAC clone screening
For historical reasons, most mouse facilities have adapted to using ES cells from CJ7/129SV strain for creating "knock-out" mice. Because
the rate of homologous recombination is affected by strain variations, it is imperative to use isogenic DNA for the construction of targeting
vector. The assembled mouse genomic sequences (NCBI, http://www.ncbi.nlm.nih.gov/genome/seq/NCBIContigInfo.html) and the
corresponding BAC clones, however, are from C57BL/6J strain, making it necessary to screen CJ7/129SV mouse genomic library.
Under BAC clone screening service, we will screen the CITB/CJ7 mouse BAC clone library
(http://informa.bio.caltech.edu/idx_www_tree.html) by PCR and provide you with up to two BAC clones as glycerol stock. The typical
turn-around time is 4 weeks. Researchers only need to provide us with cDNA accession number, and we will design PCR probes and isolate
BAC clones. In case no BAC was found, a small set-up fee is charged to recuperate the cost.
If your mouse facility offer targeting service in C57BL/6J ES cells, you can omit this step and order BAC by searching clone ID directly from
NCBI server.
This service covers BAC DNA isolation and sequencing runs from each end of the BAC clones . A gross map of intron/exon and common
restriction sites will also be provided.
Starting from a BAC clone, we will work closely with you to design a targeting strategy and make the corresponding targeting constructs. We
have a number of targeting vectors available for you to choose from and also accept researcher supplied vectors. We design and make
either standard or tissue-specific "Knock-out" constructs, or "knock-in" constructs, tailoring to the specific need of your research. Send your
inquiries of our service and pricing to KOserv@cellogenetics.com
PCR primer pairs for screening homologous recombined ES cells as well as future mouse tail genotyping will be designed and synthesized.
A plasmid clone containing the DNA fragment to be used as Southern probe for the targeted locus will be designed and generated.
The final "KO" construct DNA will be isolated under endotoxin-free condition
