云斑尖塘鳢AFLP分析技术的建立  

AFLP Molecular Marker Technology System of Oxyeleotris marmorata

作　　者：朱晓平[1] 骆剑[1] 王茜[1] 尹绍武[2] 

机构地区：[1]海南大学海洋学院海南大学热带生物资源教育部重点实验室,海南海口570228 [2]南京师范大学生命科学学院,江苏南京210046

出　　处：《激光生物学报》2012年第3期283-288,共6页Acta Laser Biology Sinica

基　　金：海南省自然科学基金项目(310037);海南省海南大学科研基金项目(kyqd1018);海南大学"211"工程研究生创新平台课题

摘　　要：以云斑尖塘鳢为对象,对其扩增片段长度多态性(AFLP)体系中的连接、预扩增、选择扩增等关键步骤的参数进行了对比实验。结果表明,在采用磷酸化接头,预扩增PCR增加延伸反应及20倍稀释预扩增产物的条件下,可得到清晰稳定的电泳图谱。所筛选的48个引物组合中,14个引物组合的产物带型稳定清晰,分布均匀,扩增片段为50-70条。本研究获得的AFLP优化参数及引物能够运用于云斑尖塘鳢群体遗传分析及分子辅助育种研究。AFLP analysis system for Oxyeleotris marmorata was established by optimizing procedures that include adapter ligation reaction, pre-amplification and selective amplifications. The results showed that quite steady results could be achieved by using phosphated-adapter, additional extension procedure before pre-amplified PCR and pre-amplified prod- ucts diluted 20 times. 14 pairs of selective primer combinations which revealed sufficient amplified bands with high reso- lution were selected from 48 pairs of primers, clear fingerprints （ 50-70 ） were obtained. Those 14 pair primers could be used to study the population genetic diversity and molecular assisted selection of O. marmorata.

关 键 词：AFLP 云斑尖塘鳢 反应体系 

分 类 号：Q31[生物学—遗传学]