卵形鲳鲹遗传多样性研究中AFLP技术优化及引物筛选

摘要

To establish an efficient and stable amplified fragment length polymorphism (AFLP) analysis system for Trachinotus ovatus, ammonium acetate method was used to extract high-quality genomic DNA. The key factors were explored and optimized for enzyme-digested reaction, ligation, elecrrophoresis and silvers staining. 10 pairs of informative primers were identified from 256 pairs of primers when new AFLP analysis system was used in the test experiment for a wild population and a cultured population of Trachinotus ovatus. The result showed that it was feasible to use the new reaction system to conduct the AFLP analysis of Trachinotus ovatus.%以卵形鲳鲹为材料,采用醋酸铵法提取高质量的基因组DNA,通过对双酶切、连接、电泳、染色等因素进行优化,建立了适用于卵形鲳鲹的AFLP反应体系.同时以卵形鲳鲹野生群体和养殖群体为材料,采用新建立AFLP反应体系从256对引物组合中筛选出10对多态性高的AFLP引物组合,获得了清晰的指纹图谱,证明建立的AF"优化体系可用于卵形鲳鲹的AFLP分析.