CAJ | 학술논문

对广西红锥种群的DNA进行提取和ISSR—PCR扩增体系进行优化。分析了退火温度、模板DNA浓度、Mg^2＋浓度、dNTPs浓度、TaqDNA聚合酶用量对反应结果的影响。红锥ISSR—PCR分析较适宜的扩增体系是：25μL PCR反应体积中，buffer（10mM Tris-HCl，pH9．0，50mM KCl，0．1％Triton X-100），1．25U Taq DNA聚合酶，4种dNTPs各O．2mM，0．4μM引物，2．0raM MgCl2，50ng模板DNA。
대엄서홍추충군적DNA진행제취화ISSR—PCR확증체계진행우화。분석료퇴화온도、모판DNA농도、Mg^2＋농도、dNTPs농도、TaqDNA취합매용량대반응결과적영향。홍추ISSR—PCR분석교괄의적확증체계시：25μL PCR반응체적중，buffer（10mM Tris-HCl，pH9．0，50mM KCl，0．1％Triton X-100），1．25U Taq DNA취합매，4충dNTPs각O．2mM，0．4μM인물，2．0raM MgCl2，50ng모판DNA。
The DNA and ISSR-PCR amplification of Guangxi Catamopsis hystrix was extracted and optimized respectively. The factors which affected the ISSR amplification such as annealing temperature, template DNA dosage, Mg^2＋ concentration, dNTPs concentration and unit of Taq DNA polymerase were selected and optimized. The results showed that the reaction system being suitable for ISSR - PCR of Castanopsis hystrix was as follows ： 1 × Taq buffer （10 raM/L Tris-HCl , pH 9.0, 50 mM/L KC1 and 0.1% Triton X-100） , 1.25 U Taq DNA polymerase , 0.2 mM 4 8 dNTP, 0.4μM primers, 2.0 mM/L MgCl2 and 50 ng template DNA in total 25 μL reaction volume.