论文部分内容阅读
比较两种从龙须菜中提取总ＤＮＡ的方法，这两种方法得到的染色体ＤＮＡ具有较好的完整性，从ＤＮＡ的产量、蛋白质含量等指标可以看出，ＣＴＡＢ法优于蛋白酶Ｋ法。实验对龙须菜ＲＡＰＤ反应条件进行了优化：在２５μＬ反应体系中，含有Ｍｇ２＋２．５ｍｍｏｌ／Ｌ，ｄＮＴＰ１００μｍｏｌ／Ｌ，总ＤＮＡ２５ｎｇ，引物０．２μｍｏｌ／Ｌ和Ｔａｑ酶１Ｕ，经９４℃变性１ｍｉｎ，３５℃退火１ｍｉｎ，７２℃延伸２ｍｉｎ，３５个循环，得到稳定的ＲＡＰＤ扩增图谱。
Comparing the two methods of extracting total DNA from Gracilaria lemaneiformis, the chromosomal DNA obtained by these two methods has good integrity. From the yield and protein content of DNA, it can be seen that the CTAB method is superior to the proteinase K method. The optimum reaction conditions were as follows: Mg2 + 2.5mmol / L, dNTP100μmol / L, total DNA 25ng, primer 0.2μmol / L and Taq enzyme 1U in 25μL reaction system, denaturation at 94 ℃ for 1min, 35 ℃ annealed 1min, 72 ℃ extended 2min, 35 cycles, to obtain a stable RAPD amplification map.