Transgenic-Specific Testing

• Copy number determination of transgenes using QPCR
• Zygosity and carrier status determination via PCR, QPCR or SNP analysis as appropriate for each model
• Expression testing of specific genes of interest, transgenes and gene knock-downs via quantitative RT-PCR (reverse transcriptase PCR)
• Microsatellite analysis for background strain characterization to ensure that client's model is on the expected strain
• Genotyping of targeted transgenics and point mutations by PCR or SNP analysis as appropriate
• Molecular phenotyping can determine transgene copy number and gene expression such as non-obese diabetes (NOD)
• Severe combined immunodeficiency (SCID) is the result of a spontaneous autosomal recessive mutation. Using PCR and restriction enzyme analysis assays we can confirm the presence or absence of this point mutation.
• Genetic Monitoring of Inbred Strains: To ensure the genetic quality and monitor the genetic drift of inbred and transgenic laboratory animals, DNA fingerprinting is available using detection of Restriction Fragment Length Polymorphisms (RFLPs), hybridization patterns, chromosome mapping, indices of band sharing, heterozygosity and genetic distance.
• Strain Characterization and Differentiation: To compare/contrast the genetic relatedness of inbred strains and to determine the presence or absence of a gene (in transgenics or knock-outs), DNA bands are compared. Homozygous animals of different strains will exhibit unique bands; heterozygotes will carry bands from both parent strains.
• Paternity Determination: DNA-based testing is performed by comparing genetic profiles of the dam, offspring and possible sires. Since the offspring inherits half the DNA pattern from each parent, eliminating the DNA known to be from the dam and comparison of remaining DNA with that of all possible sires allows almost certain identification of the actual sire. This method requires only a few drops of blood, a small piece of tissue (such as the tip of a tail or tissue from ear punch) or a swab of the mouth.
• Knockout Verification: To clarify a possible mix-up between knockout and control animals, several optimized (PCR and QPCR) assays for the most common elements (listed below) are available:
  • Neomycin
  • GFP
  • LacZ
  • Cre
  • Luciferase