【摘要】 从海带抱子体的酶解单细胞中提取的基因组ＤＮＡ，可用于随机扩增多态ＤＮＡ（ＲＡＰＤ）的重复性研究。通过对扩增反应体系中各因子和扩增程序的梯度实验，确定优化反应体系为：０．２μｍｏｌ／Ｌ引物，１．５ｍｍｏｌ／Ｌｍｇ，１００μｍｏｌ／Ｌ　ｄＮＴＰ，１.０　Ｕ Ｔａｑ ＤＮＡ聚合酶，３０ｎｇ模板 ＤＮＡ；优化反应程序为：９４℃变性２ｍｉｎ，接着 ４５个循环包括９４℃变性１ｍｉｎ，３７℃退火１ｍｉｎ，７２℃延伸２ｍｉｎ；最后７２℃延伸１０ｍｉｎ。在此优化条件下得到的ＲＡＰＤ图谱有较好的特异性和重复性，为海带遗传多样性。种质鉴定、分子标记及辅助育种等研究提供了有用的手段。更多还原

【Abstract】 Unicells isolated from Laminaria japonica sporophyte with enzymatic dissociation, were used to extract genomic DNA and to study the reproducibility of the random amplified polymorphic DNA (RAPD) patterns. The ingredients and programs of amplified reactions were individually optimized. The reactions were composed of 0.2/μmol/L primer, 1.5rnmol/L Mg2* , 100μmol/L dNTP, 1.0 U Taq DNA polymerase,30 ng template DNA. The optimized programs were predenaturation at 94℃for 2 min;and then 45 cycles containing denaturation 1 min at 94℃, annealing 1 min at 37℃ ,and elongation 2 min at 72℃; and at last holding 10 min at 72℃. The optimized conditions could result in satisfactory specificity and reproducibility of RAPD amplification patterns, and consequently provide a useful means for the research of genetic diversity, molecular marker, germplasm identification and auxiliary breeding in L. japonica.更多还原