摘要
应用多重PCR简单重复序列 (SSR)荧光标记分析技术和常规的SSR银染技术 ,对北方冬麦区 4 5 1份材料 (中国小麦初选核心种质的一部分 )进行分析 ,用相同材料和引物 ,对这两种方法的检测效果进行评价。用 2 4对引物扩增 ,银染法共检测出 2 35个等位变异 ,每个位点检测到的等位变异为 3～ 2 0 ,平均为 9 8个 ,多态性信息指数PIC为 0 2 2～ 0 93,平均 0 74 ;而荧光标记可检测到 312个等位变异 ,每个位点检测到的等位变异为 4～ 2 4 ,平均为 13 0个 ,PIC为 0 32～0 97,平均 0 75。荧光技术较银染方法在每个位点上多检测到 3个等位变异 ,检测效果更为理想 ,更适于进行遗传多样性分析和研究 ;对两种方法的费用和工作效率做了初步分析 ,表明完成 5 0 0 0× 78个反应在费用基本持平的情况下 ,微卫星荧光标记技术的检测效率显著高于银染法 (7 8倍 )。同时对微卫星荧光标记技术中低成本、高通量多重PCR体系的建立及Genescan 3 7、Genotyper 3 7软件数据分析中出现的一些具体问题进行了探讨。
Four-hundred and fifty-one wheat cultivars from Northern Winter Wheat Region were analyzed by fluorescence based genescan with multiple PCR microsatellite markers and SSR marker technology based on silver staining. In the same cultivars and the same set of SSR primers, the detection efficiency of the two systems was compared. At 24 loci among 451 accessions, total 235 alleles were obtained and average 9.8 alleles (from 3 to 20) were detected for every pairs of primers with silver staining system. However, average 13.0 alleles were detected by fluorescent system. Three more alleles were detected averagely by fluorescent system than silver-staining at one locus. With almost equal cost, the efficiency of fluorescent system was 7.8 times of silver staining system without consideration of instrument investment. Some other strategies, such as building low-cost and high-throughout multiple PCR system and data analysis using Genescan and Genotyper procedure were also discussed.
出处
《作物学报》
CAS
CSCD
北大核心
2005年第2期144-149,共6页
Acta Agronomica Sinica
基金
国家重点基础研究项目 (G19980 10 2 0 2 )
