The ability to detect multiple genetic markers in a single tube or reaction is of great importance. It is much easier and cost effective to run one reaction that detects 20 markers rather than 20 separate reactions. The advantages of multiplexing an assay are a reduction in the amount of sample consumed (important for forensic applications), a reduction in the amount of enzyme/reagents consumed (cost reduction), the collection of more information per unit time, and a simplification of data analysis. However, there are challenges in designing a multiplexed assay. The greater number of components required in a multiplexed assay can result in an increased likelihood of side reactions or failure in detecting certain markers. We are primarily concerned with multiplexing the polymerase chain reaction (PCR). In the PCR, specific regions of DNA located in a genome are amplified. The amplification process allows for detection by current instrumental techniques such as capillary electrophoresis, mass spectrometry, and HPLC. Genetic markers such as single nucleotide polymorphisms and short tandem repeats can be genotyped by the PCR and utilized for genome mapping, disease association studies, and forensic identification purposes. We are also developing tools such as primer screening software and multiplex assay condition guidelines in order to design assays with a greater success rate and in a shorter period of time.
Capillary electrophoresis; DNA; Forensics; Genotyping; Mass spectrometry; Multiplex PCR; Real time PCR; Short tandem repeat; Single nucleotide polymorphism;