ANALYSIS OF 2 MULTIPLEXED SHORT TANDEM REPEAT SYSTEMS USING CAPILLARYELECTROPHORESIS WITH MULTIWAVELENGTH FLUORESCENCE DETECTION

A series of experiments was performed to analyze the utility of capill ary electrophoresis (CE) with multiwavelength detection capabilities f or multiplex typing of short tandem repeat loci. Characteristics of th e sieving polymer, hydroxyethylcellulose, which affect resolution of s ingle strand (ss) DNA fragments were examined. Additionally, the effec ts of denaturant in the polymer system, separation voltage, and analys is temperature were studied to ascertain their effects on DNA separati ons and capillary lifetime. The use of elevated run temperature (60 de grees C) was found to improve sizing precision, to increase the lifeti me of capillaries (100 runs or more per capillary), and to provide run times of under 20 min. Finally, 100 individual human DNA samples were typed successfully using CE. The average resolution obtained was 1.4 b ases for a 200 base fragment with a standard deviation of sizing of 0. 2 bases, allowing all alleles examined to be distinguished clearly.