Pronuclear Injection

This procedure comprises three days of injections of one DNA vector into fertilized C57Bl/6J oocytes and transfer of injected zygotes into pseudopregnant females. Demonstration of a successful screening strategy to identify transgenic founders is required prior to injections. Payment of this fee does not guarantee the generation of transgenic founders because of the variability in construct integration into the genome.

 

General Overview of the Process

  1. Consult with core facility director regarding construct design.
  2. Generate construct and develop screening assay to screen for transgenic founders.
  3. Purify construct and provide documentation of optimized screening assay.
  4. Core generates pronuclear-stage embryos, microinjects DNA transgene, and transfers zygotes into pseudopregnant female mice.
  5. Core provides investigator with tissue samples to genotype pups.
  6. Investigator genotypes pups.
  7. Transgenic founders are transferred to the investigator.
  8. Founders are bred to create a transgenic line.
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