摘要
油菜SSR检测是其主要农艺性状分子标记及标记辅助育种的一项重要技术。以无花瓣品系APL0 1、常规有花瓣抗菌核病品种M 0 83及其回交群体BC1为供试材料 ,研究油菜SSR检测体系中PCR扩增的各主要成分对检测结果的影响 ,比较不同电泳方法及染色方法的效果 ,进一步优化了适用于油菜的SSR检测体系。优化后的PCR反应体系为 :1×Buffer(含 2 0 0mmol/LMgCl2 )、5 0 0 0 μmol/LdNTPs、0 5 0UTaq酶、0 0 8μmol/LSSR引物、6 0 0ng/μl模板DNA ,加ddH2 O至 10 0 0 μl。
SSR detection is an important technique of molecular marker for main agronomical traits and marker-(assisted) breeding in rape seed.An apetalous rape seed line APL01,a conventional petalous variety M083 and its backcross population were used as test materials.Effect of major component of PCR amplification on results detected in SSR detection system was studied,different electrophoresis and staining methods were compared,and SSR detection system suitable for rapeseed was established and optimized.The optimized PCR amplification system was 1×Buffer (including 2.00 mmol/L MgCl_2),50.00 μmol/L dNTPs,0.50 U Taq enzyme,0.08 μmol/L SSR primer,6.00 ng/μl template DNA,add ddH_2O to terminal volume 10.00 μl.The effect of polyacrylamide gel electrophoresis was better than that of agarose gel electrophoresis,and silver staining was better than EB staining.
出处
《江苏农业学报》
CSCD
2004年第4期225-229,共5页
Jiangsu Journal of Agricultural Sciences
基金
国家自然科学基金项目 ( 3 0 170 5 83 )
江苏省高技术项目(BG2 0 0 2 3 0 3 )
