关键词: 柱式胶回收试剂盒;DNA/分离和提纯

    摘 要:目的 确定提高柱式胶回收试剂盒DNA回收效率的最佳条件. 方法 回收的基本步骤遵循试剂盒附带的厂家说明，在此基础上，改变其中所用试剂的剂量，或者添加原试剂盒中没有的试剂，并在不同条件组合下分别进行DNA回收.电泳后，通过比较条带的亮度来比较不同条件下DNA回收效率的大小，逐步确定最佳的胶回收条件. 结果 提高柱式胶回收试剂盒DNA回收效率的最佳条件为:切胶要尽量的小;每l g胶用600μL的S1;溶胶时间:55℃，10min;沉淀时加入3/5总体积的异丙醇和1/10总体积的乙酸钠(3mol L-1 ，pH5.2);沉淀时间20min;沉淀温度为25℃(室温);用T1液溶解效率高于用水溶;溶解时间为20min(55℃).这些最佳条件中，大部分都对原来的条件进行了改良. 结论 应用了改良后的胶回收条件后，胶回收的效率提高到原来的3～4倍.

    Identification of high DNA extraction efficiency from agarose gels using gel extraction kit

    SU Ming，DOU Ke-Feng，ZHAO Ai-Zhi，LI Fu-Yang

    Department of Hepatobiliary Surgery，Xijing Hospi-tal，Fourth Military Medical University，Xi'an710033，China

    Keywords:gel extraction kit;DNA/isolation&purification

    Abstract:AIM To determine the optimal conditions for high DNA extraction efficiency from gels with gel extraction kit.METHODS The basic procedure of isolation of DNA from agarose gels were carried out essentially as described by the manufacturer.We changed the dosage of several reagents or added new reagent that was not included in the kit previ-ously and we made several different combining of these reagents.Then after electrophoresis ......