扩增片断长度多态性(AFLP)荧光标记和银染技术的比较分析

实验采用AFLP的荧光标记分析技术和常规的AFLP银染技术,对施用农药和未施用农药的狼蛛基因多态性进行分析.用4对引物扩增和银染法共检测出32条带,荧光标记技术共检测出223条带.荧光技术比银染方法在每个位点上多检测到24条带,检测效果更为理想,更适于进行遗传多样性分析和研究;对两种方法的费用和工作效率做了初步分析,AFLP荧光标记技术的检测效率是银染法的690倍,同时对AFLP荧光标记技术中低成本、高通量多重PCR体系的建立及Genescan软件数据分析中出现的一些具体问题进行了探讨.

Comparison of Fluorescence and Silver-staining Detection Systems of AFLP

AFLP fluorescence tag analyses technique and routine AFLP Silver-staining were used in this experiment.Through amplification of seven pair primers,we got 32 bands by Silver-staining technique and 223 bands by fluorescence tag technique.The result showed that,compared Silver-staining technique,we could get much more bands in every site and better checked-effect by fluorescence tag technique.In addition,analyzing or studying descendiblity polymorphism,fluorescence tag technique was more suitable than Silver-staining technique.Analyzed the cost and work efficiency,we knew that with almost equal cost,the efficiency of fluorescent system was 690 times that of silver staining system without consideration of instrument investment.Some other strategies,such as building low-cost and high-throughout multiple PCR system and data analysis using Genescanan procedure were also discussed.