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中国农学通报 ›› 2015, Vol. 31 ›› Issue (12): 205-209.doi: 10.11924/j.issn.1000-6850.2014-2456

所属专题: 油料作物

• 生物技术科学 • 上一篇    下一篇

油葵SSR-PCR反应体系的优化及引物筛选

房冬梅,吕 品,侯建华   

  1. (内蒙古农业大学农学院,呼和浩特 010019)
  • 收稿日期:2014-09-10 修回日期:2014-09-10 接受日期:2014-11-05 出版日期:2015-05-06 发布日期:2015-05-06
  • 通讯作者: 侯建华
  • 基金资助:
    内蒙古自然科学基金重点项目“油用向日葵抗旱性相关性状的QTL分析研究”(2010zd06);国家自然科学基金项目“向日葵抗(耐)旱相关性状基因QTL定位及分析”(31160288)。

Optimization of SSR-PCR Reaction System and Primer Screening in Oil Sunflower

Fang Dongmei, Lv Pin, Hou Jianhua   

  1. (College of Agronomy, Inner Mongolia Agricultural University, Huhhot 010019)
  • Received:2014-09-10 Revised:2014-09-10 Accepted:2014-11-05 Online:2015-05-06 Published:2015-05-06

摘要: 为了获得适合油葵的SSR-PCR反应体系,通过采用控制变量法对影响SSR反应体系的主要因素进行优化,研究Mg2+浓度、dNTPs浓度、Taq酶浓度、引物浓度、退火温度等因素。结果表明各因素的终浓度分别在Mg2+浓度2.5 mmol/L、dNTPs浓度0.2 mmol/L、Taq酶量0.5 U、引物浓度0.4 μmol/L及退火温度为53.9℃时最为适合。通过试验优化后的体系筛选引物,最终从50对引物筛得19对多态性好的引物可进行下一步的研究。

关键词: 芝麻, 芝麻, 施肥水平, 品种, 干物质积累与分配

Abstract: In order to obtain the SSR-PCR reaction system for oil sunflower, the method of control variable had been used to optimize the main factors influencing the SSR reaction system, which include the concentration of Mg2 , dNTPs, Taq enzyme, primer, and annealing temperature. Results showed that the optimized SSR reaction system was Mg2 2.5 mmol/L, dNTPs 0.2 mmol/L, Taq polymerase 0.5 U, primer 0.4 μmol/L and the annealing temperature 53.9℃. Through the testing system after optimization, 19 pairs of polymorphic good primer could be used for the next research from 50 pairs of primers screening.