SERVICES

Brief descriptions of all core services are listed below. Please note that several new services have been added for FY08, so if you would like additional information regarding these offerings, please contact the Core Director. All service guarantees and policies regarding project completion are underlined. Please see our Instructions section, or click on the links below for detailed information concerning sample submission and project initiation.

TRANSGENESIS:

DNA Purification/Transgenics:
Instructions   prices   order forms
The core now requires that all standard plasmid constructs be purified by the core facility prior to pronuclear microinjection for transgenic production. We request that the PI provide 50ug of purified DNA to the core, along with a plasmid map clearly indicating the relevant restriction enzyme sites. The core will then digest the DNA, gel purify it, and dilute it for microinjection. BAC constructs should be purified by the PI, and provided to the core at a minimum concentration of 10ng/ul

Transgenic Production (All strains):
Instructions   prices   order forms (B6) (FVB)
The core provides pronuclear microinjection in two inbred mouse strains, C57BL/6 and FVB, other strains can be injected upon request. The core will transfer 200 microinjected embryos into recipient animals for birth of potential founder lines. A guarantee of two positive founder lines is provided.

GENE TARGETING:

ES Cell Targeting/confirmation of HR clones:
Instructions   prices    order forms
The core will electroporate the targeting construct DNA into our stock ES cell line, HZ2.2 (129SvEv/TAC) and pick 200 drug resistant clones for expansion (neomycin, or puromycin selection cassettes). Ethanol precipitated DNA (in 96 well format) will be provided for analysis to identify recombinant clones. All targeted clones will be expanded, and a second DNA aliquot provided for confirmation. Homologous recombination at a given genetic locus cannot be guaranteed. However, our recombination frequencies are generally quite high (link to data)

ES Cell Cassette Deletion:
Instructions   prices    order forms
This service allows for the in vitro deletion of drug selection cassettes (or other target sequences) utilizing either cre or frt recombinase. Fifty clones are picked, and DNA provided for analysis. A minimum of two properly deleted clones are guaranteed.

Production of Knock-out/knock-in mice:
Instructions   prices    order forms
Correctly targeted and confirmed ES cell clones generated within the core are injected into blastocysts to generate chimeric males. Up to three independent clones are injected for each project. All high percentage chimeric male mice are bred to wild-type females and agouti offspring are produced. Germline transmission is guaranteed for all projects utilizing ES cells targeted and expanded by the core facility.

Production of Chimerics (ES cells from PI):
Instructions   prices   order forms
This service is aimed to generate chimeric mice from ES cell clones provided to the core from outside sources. Clones can either come from a) PI laboratories, b) external investigators, or c) commercial sources such as for gene-trap clones. The  core will expand a single clone, perform pathogen screening analysis (required), and
generate chimeric mice from a single day of blastocyst injections (minimum 25 blastocysts transferred). The core offers no guarantee for number of offspring or percentage of chimerism in resulting pups, nor does it guarantee germline transmission.

AUXILIARY SERVICES:

Colony Maintenance:
Contact Core Director for more information.
Colony maintenance services are charged on a per hour basis, and reflect labor fees in maintaining mouse lines for investigators on an ongoing basis. Procedures and activities included under this rate include, but are not necessarily limited to, daily monitoring and record keeping, setting up matings, weaning mice, and providing reports to investigators.

Contract Services:
Contact Core Director for more information.
This includes a wide array of services that the core can provide for investigators on an "as needed basis". All services in this category are billed on a per/hour basis. Examples of commonly provided services include, training of PI staff in animal husbandry and other techniques, necropsy and tissue collection, formalin fixation of tissues for histology, timed matings for embryo collection, and provision of reagent and tools requiring preparation by TMF staff.

Cryopreservation:
Instructions   prices  order forms
Valuable mouse lines are cryopreserved as morula stage embryos. The core will produce embryos from up to 10 mating pairs of the desired line, and freeze all embryos that are collected. A single straw will be thawed per project and tested for viability. Fifteen embryos are frozen per straw, and can be stored by the core at no additional cost.

PCR Screening:
Instructions   prices   order forms
The core provides PCR screening of genetic mouse models on a per sample basis both for animal models maintained within the core, and for those maintained elsewhere on campus. Samples for analysis can be provided as tail snips, or purified genomic DNA

Southern Blotting:
Instructions   prices   order forms
Southern blot analysis for ES cell targeting projects performed through the core is provided. Note: To utilize this service, the PI must meet with the core facility staff prior to generation of the construct for consultation on construct design, and screening strategy. Once contracted, the core will perform control Southern blot analysis utilizing wild-type DNA prior to initiation of the targeting experiment. No targeting experiments will be performed by the core until appropriate screening results for controls are obtained.

Tail Biopsy DNA Purification:
Contact Core Director for more information.
DNA is purified from tail biopsy (or other samples) as requested, and can be used for PCR or Southern blotting protocols.