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Reaction conditions were as follows for a 50 μl PCR reaction : Perkin-Elmer Cetus PCR buffer; 1.5 mM MgCl 2 , 0.25 mM dNTPs, 1.5 μl of each primer (10 μM), 1% DMSO, 5 U of Taq polymerase (Life Technologies).
In general, PCR reactions were performed in a volume of 50 μl with 50 picomoles of each primer, dNTP's (Amersham Pharmacia Biotech, Inc., Piscataway, NJ) at a final concentration of 0.5 mM, 1 μl of template, 1X PCR buffer (10X PCR buffer contained 700 mM Tris pH 8.8, 20 mM MgCl 2 , 1% Triton X-100, and 0.1% Tween 20) and Taq polymerase (Perkin-Elmer Inc., Wellesley MA) diluted to the manufacturers recommendations.
Purified PCR products were sequenced using an ABI PRISM 377 (Perkin-Elmer, Norwalk, CT) through the Oregon State University Center for Gene Research and Biotechnology.
3), 5 mM KCl, 1.5 mM MgCl 2 , 200 μM of each dNTP, and 2.5 units of Amplitaq (Perkin-Elmer/Cetus).
PCRs were performed in a model 9600 Thermocycler (Perkin-Elmer Cetus) using the following cycling protocol: initial denaturation at 94°C for 10 min; 94.