摘要
本试验将锚定PCR技术用于谷子微卫星富集文库的筛选,结果发现该技术能有效减少假阳性的产生,假阳性率仅为7.7%,该技术还能够最大限度淘汰侧翼无法设计引物的无效微卫星克隆,本研究无效克隆比率仅为3.8%。用锚定PCR技术最终筛选开发出13对多态性微卫星标记,这些标记在4个谷子材料和1个野生种中扩增出等位变异数为2~4。锚定PCR技术与常用的同位素杂交筛选技术及普通PCR技术相比,更能降低假阳性克隆和无效微卫星克隆的产生,是一种理想的微卫星文库筛选技术。
Though microsatellite markers are at present the main tool for diversity study, genetic map construction, QTL locating and marker assistant selection breeding in plants, the process for microsatellite isolation consumed considerable manpower, material resources and money. So it is very important to increase its exploiting efficiency. In this study, the anchored PCR technology was used to filter microsatellite enriched library of foxtail millet. The results showed that anchored PCR could effectively ~educe false positive clones, with a false positive ratio of 7.7 % only. Furthermore, the anchored PCR could at maximum wash out invalid microsatellite clones lacking flank sequence for primer design, with an invalid microsatellite clones ratio of 3.8% only. 13 polymorphie microsatellite markers were developed by the an- chored PCR, and the allele number ranged between 2 and 4 for these 13 polymorphic microsatellite markers when amplied in four foxtail millet cultivars and one wild relative species of foxtail millet. Compared with isotopes hybridizing and conventional PCR technologies, the anchored PCR could more effectively reduce false positive clones and invalid mierosatellite clones. It is an ideal tool for microsatellite library filter.
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2009年第4期364-367,372,共5页
Journal of Jilin Agricultural University
基金
国家自然科学基金项目(30571168)
IAEA资助项目(10757/R0)
关键词
锚定PCR
微卫星
文库筛选
anchored PCR
microsatellite
library filter
