Genotyping and Real-Time PCR Services
Genotyping; real-time PCR (RT-PCR)
This Core of the Genomics Facility performs cost-effective mouse genotyping services and qRT-PCR. For genotyping, the Core performs DNA extraction from mouse tails and toe clips and develops, optimizes and performs allele-specific PCR assays for the genotyping of knockout, knock-in, and transgenic mouse strains. This service is carried out in collaboration with the Laboratory Animal Facility (LAF) with communication through the on-line mouse genotyping database that tracks the matings, genotyping results and animal inventory for each strain of mice. The Core also offers HLA genotyping service for human samples.
Quantitative RT-PCR is offered for various purposes, including validation of DNA array gene expression data, quantification of gene expression, quantification of virus load, zygosity determination, quantitative analysis of alternative spliced transcripts, quantification of gene-specific DNA in immunoprecipitated ChIP assays, and validation of the effectiveness of siRNA transcription silencing. The Core is equipped with several complementary technology platforms that allow for comprehensive and cost-effective analysis of variations and alterations such as SNPs, mutations, gene deletions, variable nucleotide tandem repeats, LOH, and CpG methylation. These platforms include pyrosequencing (PSQ96MA, Biotage), capillary electrophoresis (CEQ8800, Beckman Coulter) and two real-time fluorescent PCR machines (ABI PRISM 7900 SDS, Applied Biosystems) in a 96-well, 384-well or low-density array format.