提高柱式胶回收试剂盒DNA回收效率的最佳条件.doc

文档介绍：提高柱式胶回收试剂盒DNA回收效率的最佳条件
作者:苏明窦科峰赵爱志李福洋
【关键词】 DNA/分离和提纯,
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,,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 ,);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.

Keywords:gel extraction kit;DNA/isolation&purification

Abstract:AIM To determine the optimal conditions for high DNA extraction efficiency from gels with gel extraction The basic procedure of isolation of DNA from agarose gels w
ere carried out essentially as described by the changed the dosage of several reagents or added new reagent that was not included in the kit previ-ously and we made several bining of these after electrophoresis,the optimal conditions for DNA extraction were determined ultimately paring the brightness of The optimal conditions for high DNA extraction from gels using gel extraction kit were:The desired fragments of DNA as small a slice of gel as possible;600μL S1solution per100mg gel;10min of dis-solving time(55℃), isopropanol and1/10vol-ume sodium acetate(3mol L-1 ,)during precipita-tion;20min of precipitation time;25