文档介绍：JIU JIANG UNIVERSITY
毕业论文
题目禺毛茛核基因的筛选
院系生命科学学院
专业生物科学
姓名
班级
指导教师
二零一二年五月
摘要
毛茛属(Ranunculus)共有2000种,我国有122种。禺毛茛复合体(R. plex)是其中形态相近的复合分类群。该复合体及其近缘种具有进化历史的悠久性、生长习性上的多样性、倍性上的多倍性以及遗传变异的丰富性等特点。SSR (Simple Sequence Repeat)已经成为一种非常成熟的分子标记技术。本文利用8对R. macranthus叶绿体SSR引物,通过SSR-PCR和变性聚丙烯酰胺凝胶电泳,在禺毛茛中筛选出2对多态性高的SSR引物(RC2F/RC2和RRC7F/RC7R)。利用这2对引物对猫爪草和肉根毛茛叶绿体基因进行扩增,扩增产物有多态性,通过这种方法可以快速鉴定猫爪草和肉根毛茛。因此,这些多态性引物的获得,为今后禺毛茛以及毛茛属植物系统进化和种间关系的研究提供更多的资料。
【关键词】禺毛茛;核基因;PCR;引物.
Abstract
Ranunculus have 2,000 species and about 122 species lie in . plex (R. plex) is posite group of taxa with similar morphology. plex and its closely related species are featured by long evolutionary history, growth habits of the diversity, ploidy and polyploidy on the richness of ic variation and others. SSR has e a very sophisticated molecular marker technology. In this paper, using eight pairs of R. macranthus chloroplast SSR primers found 2 highly polymorphic SSR primers (RC2F/RC2R and RC7F/RC7R) by SSR-PCR and denaturing polyacrylamide gel electrophoresis in R. cantoniensis. R. ternatus Thunb and R. polii Franch. ex Hemsl chloroplast genes were amplified using this two pairs of primers, PCR products had polymorphism. In this way, Thunb and R. polii Franch. ex Hemsl were identified quickly. The acquisition of these polymorphism primers will provide more information for ic evolution and this relation between species research of Ranunculus cantoniensis.
[Key words] Ranunculus cantoniensis; SSR molecular marker; Polymorphism; Primers; Screening
目录
摘要 I
Abstract i
引言 1
1
1
1
SSR分子标记介绍及研究进展 1
PCR技术 2
琼脂糖凝胶电泳 2
研究背景及意义 3
2 研究材料与实验方法 4
4
实验仪器及试剂 5
5
实验试剂 5
5
实验方法 6
总DNA 的提取步骤 6
1%的琼脂糖凝胶电泳检测所提总DNA的纯度 7
DNA与引物稀释 7
PCR扩增体系的建立 8
PCR 产物电泳检测 8
引物筛选 10
3 实验结果与分析 11
禺毛茛总DNA提取结果 11
SSR-PCR扩增结果 11
引物筛选结果 12
4 讨论 14
DNA的提取