University of Virginia Health System

Diabetes Center Mouse Genetics Core Laboratory

MR-4 room 5115

Goals:

To support the efficient development, maintenance, and expansion of genetically determined mouse models for diabetes and endocrinology research.

Objectives:

1. To provide efficient and cost-effective PCR-based genotyping for breeding colonies of mice or rats carrying targeted mutations and transgenes.

2. To use high-throughput genetic scanning via polymorphic mapped microsatellites and RFLP analysis for rapid introgression of target loci onto selected inbred backgrounds using a "marker-assisted selection" or "speed congenic" protocol in mice.

3. Use high-throughput genetic scanning via polymorphic mapped microsatellites and RFLP analysis for
 - linkage analysis 
 - characterization of genetic background
 - screening of large genomic DNA fragments in mice.

Services:

DNA isolation
High-volume PCR capability
Large library of microsatellite primers
Assay for neomycin-bearing cassettes
Expertise in “speed congenic” derivations
Expertise in linkage analysis and mapping
For investigators new to mouse breeding:
 - Advice on propagation and maintenance of mouse lines
 - Education in database management
 - Sample IACUC protocols

Single-Allele Genotyping:

High-volume specialized genotyping (>50 samples/month) with DNA isolation
 and 48-hour turnaround for scheduled samples
Determination of zygosity via flanking microsatellite analysis
 Can replace 3-primer PCR assays and Southern blots
Consultation for development of PCR-based allele-specific assays
Development of “multiple mutant” strains

Breeding Techniques:

Model IACUC protocols
Principles of mouse husbandry
Breeding strategies and identification
Record-keeping, including a simple-to-use MS Access database for tracking lineage, genetic, and phenotypic data
Protocols for tissue sampling
Instruction in the use of genomic databases

This new core consists of a mouse breeding facility and a genotyping facility.  The breeding facility provides the following:  an efficient and controlled unit for propagation and selection of mutant genes or specified genetic traits in mice; tissue sampling and phenotypic assessment of each animal based on protocols established for each project; and tracking of each animal individually using a database designed to store phenotypic, genotypic and breeding data in a searchable format.  The genotyping facility provides DNA isolation and PCR-based mutation or linkage analysis using protocols designed in consultation with each investigator.  The Director, technicians, and consultants assembled for this core are seasoned investigators with specific technical expertise and experience in the development of new techniques. 

The Mouse Genetics Core was initially established in 2001 with a Pratt commitment of $50,000. The original goal of this core was to provide expertise and facilities for the manipulation of the genetic background of mice that had been genetically engineered through the University’s transgenic and knock-out mouse facility, or had been provided to University investigators from other sources. This initiative represented an expansion of an activity that has functioned for several years as a portion of an National Institutes of Health (NIH)-sponsored program project in systemic lupus erythematosus. The Core Facility interfaces closely with the Transgenic Mouse Core and the Biomolecular Resource Core.