Development of microsatellite markers for radish breeding

萝卜育种微卫星标记的开发

• 批准号: 11556002
• 负责人: FUJIMURA Tatsuhito
• 金额: $ 8.64万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11556002/
• 关键词: genome; microsatellites; genetic map; ダイコン; マグネチックビーズ; 遺伝子マーカー; ダイレクトシークエンス; 育種

This research was planed to develop the new method for constructing the microsatellite markers of radish which will form a back born of radish genome map. Those makers will facilitate radish breeding. Radish DNA was digested with two kind of restriction enzymes and then ligated with two kinds of adapters specific to each restriction sites. These chimeric DNA fragments were preliminary amplified by PCR, and then hybridized to DNA oligomer composed of microsatellite motif on a nylon membrane, DNA fragments hybridized were eluted from a membrane, and amplified with selective PCR printers with one to two base 3' extensions. Reduced number of PCR products were separated easily on an acryl amide gel electrophoresis. These separated PCR products were sequence directly and finally constructed twenty microsatellite markers.

本研究旨在开发构建萝卜微卫星标记的新方法，形成萝卜基因组图谱的后生。这些制造商将促进萝卜育种。用两种限制性酶消化萝卜 DNA，然后与每种限制性位点特异的两种接头连接。这些嵌合DNA片段通过PCR初步扩增，然后与尼龙膜上的由微卫星基序组成的DNA寡聚物杂交，杂交的DNA片段从膜上洗脱，并用选择性PCR打印机扩增，具有1至2个碱基3'延伸。通过丙烯酰胺凝胶电泳可以轻松分离数量减少的 PCR 产物。将分离的PCR产物直接测序，最终构建20个微卫星标记。