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 Transgenic Mouse Facility |
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UCI > Research > TMF > Summary of Services Summary of Services and PricesThe TMF currently offers the following services to investigators at UCI. Some services are also available to other UC campuses and to clients outside of the UC system. Note: C57BL/6NHsd is the Harlan substrain of C57BL/6, C57BL/6NTac is the Taconic substrain, and C57BL/6J is the Jackson Laboratory substrain. These substrains are not genetically identical. CB6F1 is a hybrid strain produced from crosses of BALB/c females with C57BL/6 males. When CB6F1 mice are used as egg donors, the eggs themselves (and the resulting transgenic founders) are CB6F2. The patent covering the method of pronuclear injection of DNA to make transgenic mice has expired, so services utilizing this method are now available to any client. Production of Transgenic Mice by DNA MicroinjectionBasic DNA microinjection service: The TMF will harvest and inject pronuclei of fertilized mouse eggs with DNA supplied by the investigator. Instructions on preparing the DNA are available. The final purification step will be done by the TMF. A single Basic Service will not always result in transgenic offspring being produced. However, the TMF does guarantee that at least 100 eggs will be implanted or at least 3 transgenic offspring produced (whichever comes first). Brief descriptions of the construct and its expected phenotype are requested. Prices vary by donor strain and client’s affiliation, as described in the following table.
*Payment must be made from a UC account. Shipping fees are extra. Premium DNA microinjection service: DNA supplied by the investigator will be injected into fertilized eggs until at least 50 pups (or embryos) or at least 3 transgenic offspring are produced, whichever comes first. Additionally, the TMF will tail-cut all offspring, identify them by ear-punch, and deliver the tail samples to the client for genotyping (via FedEx on dry ice for off-campus clients). Transgenic pups identified by the client are shipped or transferred after weaning. Transgenic status refers only to DNA integration. The TMF cannot guarantee expression of RNA or protein from the integrated DNA. Instructions on preparing the DNA are available. The final purification step will be done by the TMF for plasmid-based transgenes. BACs and YACs must be purified entirely by the client. Brief descriptions of the construct and its expected phenotype are required. Prices vary by donor strain and client’s affiliation, as described in the following table.
*Payment must be made from a UC account. Shipping fees are extra. PRODUCTION OF CHIMERIC MICE BY INJECTION OF TARGETED ES CELLS INTO BLASTOCYSTSES cell injection service: The TMF will harvest blastocysts from C57BL/6NTac mice and inject 10-15 ES cells, provided by the investigator, into each viable blastocyst. Injected blastocysts will then be implanted into pseudopregnant females. The TMF guarantees that at least 50 blastocysts will be successfully injected and implanted into foster moms, or 3 chimeric pups will be produced, whichever comes first. Testing ES cells for pathogens: ES cells supplied by the client (i.e., not produced by the TMF itself) must be tested for mycoplasma and mouse pathogens before injection. Clients must submit one extra vial of frozen cells for this purpose. Testing fee: $200. Expansion of ES cell clones to freeze back extra vials: We recommend this service in cases where the client has obtained a single vial of cells, e.g., from BayGenomics or some other library, and wants to have some back-up vials frozen down, in addition to having the cells injected. The price varies according to the type of ES cells. Generally, about 8-10 vials are frozen back, depending on the growth characteristics.
*Payment must be made from a UC account. Shipping fees are extra. PRODUCTION AND CHARACTERIZATION OF TARGETED ES CELLSES cell electroporation, clone picking, and DNA extraction: Using a targeting construct supplied by the client, we will electroporate the construct into embryonic stem (ES) cells, grow the cells under selection, pick up to 384 colonies, split each colony into twin sets of plates, freeze one set, and extract DNA from the other set. We strongly urge clients to consult with us regarding the design of their targeting constructs and alternative methods of obtaining targeted ES cells. Genotyping of ES cell colonies by Southern blot: We will screen the colonies from an ES cell electroporation to determine which ones have undergone homologous recombination. The client must supply a DNA probe that has been tested on a Southern blot of genomic DNA to prove it detects the correct wildtype band size. Note: purchase price of restriction enzyme is extra. Expansion of selected ES cell colonies from 96-well plates: We will thaw colonies designated by the client, expand them, freeze down ~8-10 tubes of cells from each colonies, and extract at least 50 ug of DNA from each colony for further analysis of the targeted allele. Karyotyping: Cells are grown, metaphase spreads prepared, and digital images of spreads are analyzed to determine the number of chromosomes per spread, for a minimum of 30 spreads. Secondary electroporation of targeted ES cell clones: We will perform a second electroporation with an expression plasmid or similar construct on one clone selected by the client. This service is intended for the transient introduction of a recombinase (Cre or Flp) to effect recombination of loxP or frt sites. Up to 192 colonies will be picked, split, frozen, and DNA extracted.
*Payment must be made from a UC account. MISCELLANEOUS SERVICESBreeding Service: The TMF will breed genetically-modified mice in its holding rooms. The cost includes tail-cutting and identification of pups, with tail biopsies supplied to the client for genotyping. A flat fee per cage per day is charged, plus the cost of any mice purchased by the TMF. Rederivation of mice by embryo transfer: The TMF will superovulate, mate, and collect embryos at the 8-cell stage, using mice supplied by the investigator. The embryos will then be implanted into foster moms prepared by the TMF. Price includes serology of foster moms. This service is used for elimination of mouse pathogens and can accommodate a maximum of 30 females per session. We require at least 4 females per strain. Cryopreservation of mouse embryos: Using both male and female mice supplied by the client, 8-cell stage embryos, harvested from superovulated and mated females, will be frozen and stored in liquid nitrogen in the TMF. The price is for one cryopreservation session, regardless of embryo yield. If sufficient embryos are available, the TMF will thaw a small portion and transfer them to foster moms to demonstrate ability to recover live mice. One session can accommodate up to 30 females. We require at least 15 females per strain. In vitro fertilization: Using mice supplied by the client, the TMF will harvest oocytes from superovulated females and fertilize them in vitro with sperm harvested from one or more males. Embryos are cultured overnight to the 2-cell stage and implanted into pseudopregnant foster moms. If no pups are obtained, the TMF will repeat the procedure once at no extra charge. The small-scale service accommodates up to 8 females. The large-scale service accommodates up to 30 females. PCR genotyping service: The TMF will perform PCR, using mouse genomic DNA, primers, and reaction conditions supplied by the investigator. The products will be separated by gel electrophoresis and photographed to document results. Each procedure can accommodate up to 24 DNA samples. DNA isolation and genotyping service: The TMF will collect tail biopsies from mouse pups and ear-punch the pups for identification. DNA will be extracted from the biopsies and analyzed by PCR, using primers and a protocol of demonstrated utility, supplied by the investigator. Each procedure can accommodate up to 24 biopsies. This procedure is primarily intended for transgenic mice being bred by the TMF for an investigator.
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