PAGE银染和条带回收方法的改进

Improvement on PAGE Silver Staining and Band Recovering

为了提高聚丙烯酰胺凝胶电泳（PAGE）的银染效果和条带回收的效率,以广玉兰DNA为材料进行ISSR-PCR分析,比较了两种银染（常规银染法和快速银染法）和条带回收法（乙醇糖原法和改进的煮沸法）.结果显示：两种银染方法效果一致,但常规银染要用2-3 h,而快速银染仅用30 m in,是常规银染的1/6-1/4;改良的煮沸法回收条带比乙醇糖原法更简便、更快捷.因此,快速银染和改良的煮沸法条带回收法可以应用在以后的研究中.

To enhance the efficiency of silver staining and band recovering in Polyacrylamide Gel Electrophoresis（PAGE）,both conventional silver staining and speedy silver staining were investigated,using the technique of ISSR-PCR markers with Magnolia grandiflora DNA.During band-recovering,conventional ethanol hepatin method and improved boiling method were employed and followed by re-amplification of PCR identification.The results indicate that the two methods produce similar clear bands and band patterns,but the procedure of conventional silver staining takes 2～3 hours while the speedy silver staining only requires 30 minutes which reduces the experimental time by 4～6 times.The improved boiling method is more convenient and time-saving than the conventional one.Speedy silver staining and improved boiling band-recovering can be employed in the future studies.