耧斗菜属AFLP体系的建立和优化

Optimization of AFLP of Aquilegia

通过对AFLP反应体系的主要的四个影响因素:Mg2+、dNTPs、Taq酶、引物进行四水平筛选试验,利用三种耧斗菜Aquilegia parviflora、A.viridiflora、A.vulgaris优化了耧斗菜属的AFLP反应体系,采用64对引物组合对三种耧斗菜进行了PCR扩增和聚丙烯酰胺电泳检测。其中9对引物组合表现出较高的稳定性、清晰度和多态性。初步建立了耧斗菜的AFLP反应体系,得到质量比较好的AFLP胶图。从而为AFLP分子标记技术应用于耧斗菜属的研究奠定坚实基础。

In this study,the design was used to optimize AFLP-PCR amplification system on Aquilegia on four levels of four factors（Mg2＋,dNTPs,Taq polymerse and primer） .The amplification was performed using 64 pairs of primer combination and the products were separated on 6% polyacrylamide gels.9 pairs of primer combination can produce scorable,clear,suitable and polymorphic bands.According to these experiments,the AFLP-PCR amplification system of Aquilegia was constucted and AFLP fingerprint was obtained.This optimized system for AFLP would become one of the protocols for further research on Aquilegia.