Shared Facilities

Transgenic & Knock-Out Mouse

Policies & Procedures

Transgenic & Knock-Out Mouse Core service charges are as follows. See below for transgenic and knock-out mouse production procedures.

Transgenic & Knock-Out Mouse Core Service Charges
(Effective July 2014)
Service Pricing
Pronuclear injection  $3,562
ES transfection $6,861
Blast injection $8,090
Single cell line binj   $3,549
Assisted reproduction
Sperm freeze $628
IVF   $1,310
Embryo freeze    $764
Sperm cryo and verifi $1,638
Thaw and transfer   $655
Cryo storage $55/line/yr.
Core Policy Changes
  1. The transfection charge is per transfection. A second transfection is a second charge.
  2. Alternative strains can be used for pronuclear or blast injections, but the user must pay all additional mouse costs, including per diems while waiting for results.
  3. If no founders are found from the first round of pronuclear injections, the user checks the DNA prep, then reinjection is done at no charge. If no founders are found from second round of injections, the user is charged for any additional injections.
Procedures
  1. For transgenic mouse production, investigators provide the DNA construct to the core. The core then:
    • Harvests single cell embryos
    • Microinjects the embryos with the DNA construct
    • Transplants the microinjected embryos into pseudo-pregnant females
    • Performs Cesarean sections as required
    • Provides the resulting pups at weaning age to the investigator for analyses
  2. For knock-out mouse production, the core provides three types of services:
    • For ES cell transfection, the investigator provides the core with a targeting vector. The core then:
      • transfects ES cells with the targeting vector
      • selects the resulting transfected clones
      • generates frozen stocks of the clones
      • provides the investigator with DNA samples from each of the clones for molecular analyses
    • For blastocyst injections, the investigator provides (or identifies) the desired ES clone. The core then:
      • recovers and amplifies the ES cell line
      • provides the investigator with cells for additional molecular and cell biologic analyses
      • generates backup frozen stocks
      • injects the cells into blastocysts
      • transplants the injected blastocysts into pseudo-pregnant females
      • performs Cesarean sections as required
      • culls the resulting chimeric animals
      • either provides the chimeric pups at weaning age to the investigator for analyses or uses the pups for rapid germ-line breeding
    • For rapid germ-line breeding, core personnel:
      • rotates female mice with the chimeric males
      • replaces plugged females until a total of 10 pregnancies is culled
      • screens the resulting pups for germ-line transmission of the ES cell genome (determined via coat color)
      • provides tail biopsies to the investigator for molecular confirmation of germ-line transmission
      • provides the resulting heterozygous animals at weaning age to the investigator for analysis