生命科学研究2009，Vol.13Issue(3)：251-257,7.
毛细管电泳四色荧光检测法分析茶树SSR标记
Tea Plant Microsatellite Genotyping by Capillary Electrophoresis with Four-Color Fluorescent Detection Technique
摘要
Abstract
The capillary electrophoresis with four-color fluorescent detection technique was used to analyze the microsatellites in tea plant. Three primers were used in one PCR reaction for amplification of a defined microsatellite locus by this method. The three primers were as follows: a sequence-specific forward primer tailed with M13-tail at its 5' end, a sequence-specific reverse primer and a universal fluorescent-labeled MI3 primer (5'-CACGACGTTGTAAAACGAC-3'). In order to detect three or more microsatellite loci simultaneously in each capillary electrophoresis through four-color fluorescent detection system, three universal M13 primers were labeled with three different fluorescence dyes (blue,green and black). With this method the genetic analysis of 16 microsatellite loci among 42 tea plants was carried out. The results showed that this method had merits of simple, reliable, cost-effective and high-throughput microsatellite genotyping, and the economy brought by it became greater with an increasing number of microsatellite markers. Also, 11 polymorphie microsatellite markers of tea plant were identified by this method, and these markers would be useful for investigating genetic questions of tea plant (Camellia sinensis ).关键词
微卫星(microsatellite/SSR)/通用M13引物/毛细管电泳/四色荧光检测/茶树Key words
microsatellite/universal M13 primer/capillary electrophoresis/four-color fluorescent分类
农业科技引用本文复制引用
黄建安,李娟,谭月萍,刘硕谦,李勤,刘仲华..毛细管电泳四色荧光检测法分析茶树SSR标记[J].生命科学研究,2009,13(3):251-257,7.基金项目
国家自然科学基金资助项目(30871572) （30871572）
湖南农业大学人才稳定基金资助项目(07WD22) Project of National Natural Science Foundation of China (30871572) （07WD22）
Hunan Agricultural University Talent Stabilization (07WD22). （07WD22）
