吉富罗非鱼AFLP反应体系的建立与优化

Establishment and Optimization of AFLP Analysis System for GIFT Tilapia

本文以我国水产养殖主导品种吉富罗非鱼(farmed tilapia)为研究材料,进行扩增片段长度多态性(AFLP)反应体系的研究,初步建立了一套适合罗非鱼的AFLP反应体系。对体系中关键环节的优化结果如下:基因组DNA要求无降解和RNA污染,OD260/OD280比值在1.8~2.0之间;基因组DNA EcoRⅠ/MseⅠ37℃双酶切6h、20℃连接20h,连接产物10倍稀释,PCR预扩增产物稀释45倍作为选择性扩增的模板,选择性扩增产物经6%变性丙烯酰胺凝胶电泳检测可获得条带清晰、背景干扰小的图像。该体系的构建为今后进行罗非鱼群体遗传多样性分析、种质鉴定、重要经济性状分子标记的开发及遗传连锁图谱构建等方面研究提供了参考。

In this article, we selected genetic improvement of farmed tilapia (GIFT), which was the major cultured fishes in China as the study material, to investigate an AFLP (amplified fragment length polymorphism) reaction system, and to initially establish a set of AFLP reaction system for tilapia. By optimizing the key factors in the system, the results showed that the quality of template DNA was no degradation and RNA pollution-free and the OD260/OD280 was 1.8~2.0. The genome DNA was digested at 37℃ for 6 h by ...