林木RAPD分析及实验条件的优化

随机扩增多态分子标记是目前林木遗传研究中使用最为广泛的一种分子标记，作者对影响ＲＡＰＤ实验的因素进行了分析和探讨，确定了一针对林木进行ＲＡＰＤ分析的实验程序。根据该程序，可以快速确定ＲＡＰＤ实验的理想条件，获得ＲＡＰＤ反应的良好重复性。

RAPD Experiment Protocol and Optimization in Forest Trees

RAPD marker is the most widely adopted molecular marker in forest genetic studies. The main reason is that this kind of markers can be obtained without any DNA sequence information about the studying material before hand. So it is especially suitable for the organism which little molecular study has been carried on. We can design primers arbitratily according to a common rule and the experiment process is relatively simple compared with other kinds of markers, such as RFLP and SSR. But this kind of marker has a fatal shortcoming, if the reaction condition is not suitable for the studying material, it gives out very flexible bands and cant be repeated. So the suitable amplification condition is critical to RAPD analysis. In this paper, the influencing factor on RAPD experiment was analyzed, such as the annealing temperature, the Mg2+ 2]concentration, the template concentration and so on. An optimized protocol especially suitable for forest trees was put forward through a series of experiments. According to the protocol, the RAPD condition can be fast determined for different material and the RAPD bands can be well repeated.