山桐子ISSR-PCR反应体系的建立与引物筛选

Establishment of ISSR-PCR reaction system of Idesia polycarpa and the screening of primers

以山桐子幼嫩叶片为材料,通过对CTAB法(十六烷基三甲基溴化铵法)的改良,摸索出适于山桐子基因组DNA提取的方法,并以核酸纯度、片断分布情况等指标来评价,获取高质量基因组DNA;用2个不同样品对65条ISSR引物进行了筛选,筛选出了22条扩增图谱清晰的ISSR引物.结果表明,获得的基因组质量较高,获得的ISSR引物适用于山桐子,同时也表明反应体系对山桐子的ISSR分析是可行的;并对影响山桐子基因组DNA的制备、引物筛选及扩增反应因素进行了简要的分析讨论.

Genomic DNA was extracted with modified CTAB method from tender leaves of Idesia polycarpa. DNA quality was evaluated by indexes like purity and distribution of nucleic acid. High-quality genomic DNA was obtained. Sixty-five ISSR primers were screened with two different samples. And twenty-two clear and stable amplification patterns were obtained. The results indicated the DNA was good and the system was feasible. The factors affecting the DNA extraction, the screening of primers and amplification of genome DNA of Idesia polycarpa were discussed in the paper.