Real-time PCR
The development of real-time detection of PCR products has proven to be a major advancement in how DNA and RNA quantitation is performed. This technology allows the detection of PCR products during the linear phase of amplification while the products are actually being generated rather than at the end of a fixed number of cycles. During this linear phase, there is a direct quantitative relationship between the amount of starting target sequence and the amount of PCR product generated. Thus, this technology provides a reliable, accurate, and rapid method to quantitate nucleic acid, which is not available through traditional PCR methods.

Capillary Electrophoresis
The ABI PRISM 3100-Avant Genetric Analyzer is a flurorescence based DNA analysis system utilizing capillary electrophoresis. It offers continuous, non-attended operation with automated polymer loading, sample injection, separation and detection, and data analysis. This 4-capillary instrument is capable of running both sequence and fragment analysis.

Ongoing projects:

• Analyzing human chromosome 1 abnormalities in renal oncocytoma by loss of heterozygosity.
  A subset of renal oncocytoma has previously been shown to lack chromosome 1 or 1p by conventional cytogenetics, and this loss is potentially associated with the loss of a tumor suppressor gene. As there is currently no "gold standard" for these types of studies, the same samples were evaluated by FISH and, subsequently, by fragment analysis. For fragment analysis, paraffin sections from tumor and adjacent normal tissue were studied by amplifying three microsattelite markers: D1S508, D1S199, and D1S2734. The results of this technique show excellent correlation with cytogenetics and FISH in tumors with loss of 1/1p, and cases of discordance may eventually lead to the delineation of a minimum deletion interval for the proposed tumor suppressor gene.
• Captive cheetahs may experience a stress response to environmental change that affects immune function.
  The captive North American cheetah (Acinonyx jubatus) is highly endangered because of loss of habitat in the wild and failure to thrive in captivity. The captive population is not self-sustaining because of high prevalences of unusual diseases and poor reproductive success. Cheetahs are commonly moved between zoos for breeding purposes to maintain genetic diversity within the captive population, but movement may exacerbate infertility and disease in this species. Using cross-species primers and capillary sequencing, novel cheetah interleukin (IL) -10 sequences have recently been obtained. Hopefully, these sequences will lead to a greater understanding of the mechanisms underlying disease development in this species, and identify risk factors that can direct disease prevention and treatment.