International Knockout Mouse Consortium - Knowledgebase

The clones labelled as "Knockout first" are expected to contain all the loxP sites as shown in the allele map. The knockout is obtained by introduction of a splice acceptor/reporter- cassette with a strong polyA site into an endogenous intron upstream of a critical exon. Should this allele be...

If the cassette has been successfully flipped out the internal LacZ PCR would be expected to fail, but the mutant-specific PCR would be expected to still amplify a band in mutant mice, as it is 5’ to the first FRT site. In homozygous mutant mice the wild type PCR would be expected...

As the WTSI MGP generates the mutant mouse lines for phenotypic analysis we are keen to offer available mice whilst on the shelf to the scientific community who have expressed an interest. However WTSI MGP is not a distribution centre that breeds for export. The mutant lines generated by WTSI...

One of the methods used in the WTSI MGP QC to confirm targeting of the intended gene by the mutation cassette is LR-PCR. WTSI MGP uses 3’ LR-PCR to confirm targeting. The primers used are a the 3’ arm FRTL Universal primer (5’ primer) and either the GR3 or GR4 gene specific 3’primer that...

SR-PCR genotyping uses a combination of separate PCR reactions that detect the presence/absence of the cassette lacZ (confirms the presence of the cassette), the gene-specific wild type allele and the gene-specific mutant allele by short range PCR.  Interpretation of the consolidated...

We use a qPCR reaction to count the neo gene marker present in the targeting cassette; heterozygotes will have one copy of the Neo gene, a homozygote will have two copies and a wild type will have no copies. Quantitative real time PCR (qPCR) copy number detection works by comparing the...

The mouse model the MGP sends will be supplied with an information sheet relating to the genetic background. This details contributing elements such as the Embryonic Stem Cell origin, Test Cross for germ line transmission and subsequent backcrossing that may have been carried out. The genetic...

In designing a strategy for the introduction of FlpeR and Cre lines consideration to the mutation type should be contemplated. Allele maps may be viewed at the HTGT website and searching for the gene in question and viewing the details .The clone ID used at microinjection to generate the colony...

Taking some Acot6 mutant mice as an example: Colony Prefix Name Genotype Coat Sex D.O.B. Age (weeks) Earclip ...

As the mutant mouse lines generated by WTSI MGP pass through the WTSI MGP primary phenotypic characterisation studies the information generated may be viewed through the WTSI Mouse Portal (www.sanger.ac.uk/mouseportal ) where the gene target in question may be searched for. A Heat Map of...