For pricing information, please check our Fees page.
Generation of transgenic mice by pronuclear injection or CRISPR/Cas9 injection
We will perform three days of injections. For most injections we use B6D2 hybrid embryos or C57BL/6 embryos. Upon request we may be able to work with other strains. Information on how to prepare your plasmid DNA for submission to the core can be found in the Forms and Instructions section. Check with the core for prepping RNAs for CRISPR injections.
Generation of chimeras by blastocyst injection (129 and 129/B6 hybrid ES cells)
We will perform three days of injections and inject up to three different ES cell clones. The core cannot guarantee the generation of chimeric mice or germline transmission of the ES cell mutation.
Generation of chimeras by blastocyst injection (C57BL/6 IKMC ES cells)
We will inject three different ES cell clones if available. We highly recommend that you purchase three clones for your gene from the International Knockout Mouse Consortium (IKMC) groups (EuCOMM, KOMP, NorCOMM and TIGM) as the germline transmission rates for IKMC C57BL/6N clones are lower than 129 or 129xB6 hybrid clones. In addition, typically more blastocysts (50-60) need to be injected for each C57BL/6N clone compared with 129 or 129xB6 hybrid ES cell clones to generate a sufficient number of high percentage male chimeras. The core cannot guarantee the generation of high percentage chimeric mice or germline transmission of the ES cell mutation.
Expansion of ES cell clones
Clones from the International Knockout Mouse Consortium will be accepted by the MGEF. All other ES cell clones will be first screened for pathogens (IMPACT SC - IDEXX BioResearch) before the MGEF will expand them. Typically we will prepare 6 vials of cells per clone.
Electroporation of gene targeting vector and selection
We will electroporate your targeting vector into ES cells and select with the appropriate drugs. Clones will be picked and expanded and then duplicate 96 well plates prepared. One plate will be frozen and the other duplicate plate will be given to the investigator for genotyping. Up to three 96 well plates will be picked for analysis.
Cryopreservation of mouse embryos
Embryos will be collected and frozen at the two-cell stage from up to 10 females per session. Please note that more than one session may be needed to freeze a sufficient number of embryos. Following freezing a test straw will be thawed and the embryos will be cultured in vitro to verify their viability.
Long-term storage of cryopreserved embryos and sperm
Embryos and sperm are stored in liquid nitrogen in two separate containers.
Recovery of cryopreserved embryos
Embryos stored in liquid nitrogen are thawed and then surgically transferred to one or more recipient female mice and the embryos are allowed to develop to term.
Rederivation of mouse lines
Embryos that contain an intact zona are isolated from donor female mice provided by the investigator, extensively washed in culture medium and then surgically transferred to a recipient female mice, which is placed in quarantine. After the resulting pups are weaned, blood from the recipient mouse is tested for pathogens. If the pathogen testing is negative, the mice can then be transferred to the investigator.
Labor and Consultation
Labor includes things like help with mouse husbandry or assistance with embryo collections. Consultation typically refers to significant intellectual effort made by MGEF personnel related to the design or implementation of a project. It does not refer to the routine discussion of a project with an investigator or their lab personnel.