Evaluation of sieving polymers for fast, reproducible electrophoretic analysis of short tandem repeats (STR) in capillaries

Efficient capillary electrophoretic STR analysis requires rapid, reproducible and robust separation of DNA fragments with reasonable capillary longevity--this is currently accomplished using proprietary commercial polymeric sieving matrices specifically developed for this separation. These matrices, while effective, are costly and do not provide adequate resolution of STR DNA fragments in capillaries with shorter effective separation lengths, increasing the time required to accomplish the separation and minimizing the potential extrapolation to other miniaturized platforms. As the forensic community looks toward next generation microchip technology as a means of processing casework more rapidly, new sieving polymers need to be evaluated for utilization in this platform. The research presented here describes the assessment of commercially-available polymeric sieving matrices for STR analysis, with consideration given to feasibility of incorporation into a microdevice. Polymer composition, molecular weight, and concentration were evaluated, along with an assessment of the effects of buffer composition, separation temperature, and capillary length. These variables were evaluated individually or collectively on the ability to resolve STR DNA fragments and the reproducibility of the separations and the results compared to a proprietary commercial product. A 600,000 Da MW poly(ethylene oxide) (PEO) solution at a 3% (w/v) concentration was determined to be the most suitable matrix for these separations. This polymer, in coated capillaries, provided highly robust and reproducible separations, with near baseline resolution of fragments having single base differences. Reductions in the temperature of the separation, from 60 degrees C to 40 degrees C, and the urea concentration of the buffer, from 7 M to 3.5 M, provided increased longevity of the PEO polymer for repeated separations. Comparison of this polymer with currently specified commercial products used for STR analysis showed that the optimized PEO matrix provided superior separations under all conditions tested. In addition, PEO could be utilized in shorter capillary systems, with a concurrent decrease in analysis time, highlighting its potential for use in shortened capillary or microdevice systems.     

U.S. SOUTH AFRICA POLICY IN THE 1980s: CONSTRUCTIVE ENGAGEMENT AND BEYOND

In this article no attempt is made to defend the Reagan administration's six-year-old policy of constructive engagement toward the Republic of South Africa, for to all but a few diehards the policy is indefensible. Instead, my response to Beres contains three components. First, I take issue with his dismissal of constructive engagement as no more than a ruse by which "to make apartheid palatable" to the American people, and argue that the purpose of the policy was to enable the Reagan administration to take credit for achieving a Namibia independence settlement. Second, I show that Beres' plea for popular opposition to an unjust policy comes too late, for a large number of Americans, during the past two years in particular, have been doing his bidding. Finally, I conclude with an assessment of the nature of the policy debate over South Africa that such popularity has prompted, in which direction U.S. South Africa policy now appears headed, and what issues, if any, remain to be resolved.