【摘要】 笔者以24份不同来源地的野生赤苍藤种质为试验材料,采用L₉（3³）正交试验对赤苍藤SCoT-PCR反应体系进行优化,并通过优化后的体系进行引物筛选,结果表明,PCR体系中各因素对多态性结果的影响程度排序为DNA模板量>引物用量>2×Taq Master Mix用量。最佳反应体系（20μL）为1 μL DNA模板(50 ng·μL^-1)、1.2 μL引物(10 μmoL·μL^-1)、10 μL 2×Taq Master Mix和7.8 μL ddH₂O。通过该体系从36条SCoT引物中筛选出14条稳定性好、多态性高的引物,并确定了最佳退火温度。该体系的建立和引物的筛选将为后续开展赤苍藤种质资源收集与保护、遗传多样性研究和分子育种提供参考依据。更多还原

【Abstract】 In this study, the SCoT-PCR reaction system of Erythropalum scandens Bl. was optimized by L₉（3³）orthogonal design,and 24 samples from different origins were used as experimental materials for primer screening and system verification. The factors affecting polymorphism were listed as follows, the amount of DNA template > the amount of primers > the amount of 2×Taq Master Mix, and the best reaction system（20 μL）was 1 μL DNA template(50 ng·μL^-1), 1.2 μL primer(10 μmoL·μL^-1), 10 μL 2×Taq Master Mix and 7.8 μL ddH₂O. Totally 14 primers with good stability and high polymorphism were selected from 36 SCoT primers, and the optimal renaturation temperature was determined. The establishment of the system and the screening of primers will provide a reference for the subsequent collection and protection of germplasm resources, genetic diversity research and molecular breeding of Erythropalum scandens.更多还原