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中国农学通报

中国农学通报 ›› 2006, Vol. 22 ›› Issue (12): 397-397.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

苏丹草RAPD反应条件的优化与应用

李杰勤,王丽华,詹秋文,李云飞   

  • 出版日期:2006-12-05 发布日期:2006-12-05

Optimization of RAPD Reaction System for Construction of the DNA Fingerprinting in Sudangrass

Li Jieqin, Wang Lihua, Zhan Qiuwen, Li Yunfei   

  • Online:2006-12-05 Published:2006-12-05

摘要： 为研究不同因素对苏丹草随机扩增多态性DNA（RAPD）反应体系的影响，建立并优化反应体系；再利用优化体系绘制了DNA指纹图谱以区分两个高丹草品种。提取苏丹草的基因组DNA，分别设置Mg2+浓度为1.0 mmol/L、1.5 mmol/L、2.0 mmol/L、2.5 mmol/L、3.0 mmol/L；退火温度为34℃、35℃、36℃、37℃、38℃；Taq酶含量为0.6U、0.8U、1.0U、1.2U、1.4U进行PCR扩增。结果表明：Mg2+2.0mmol/L，退火温度36度，Tap酶活性值1.2U是进行苏丹草RAPD分析的最优反应条件。利用最优反应条件绘制了皖草2号和皖草3号的DNA指纹图谱，在该图谱中有A、B两条特征带，可以区分皖草2号和皖草3号。

Abstract: To establish and optimize reaction system, the effects of different factors on random amplified polymorphic DNA (RAPD) reaction system in Sudangrass were studied and the fingerprinting of two hybrids to differentiate them were constructed. Genome DNA was isolated. The different concentration of Mg2+(1.0mmol/L, 1.5 mmol/L, 2.0 mmol/L, 2.5 mmol/L, 3.0 mmol/L), annealing temperature (34℃, 35℃, 36℃, 37℃, 38℃) and Tap DNA polymerase activity (0.6U, 0.8U, 1.0U, 1.2U, 1.4U) were designed for PCR amplification. The results were as follow: the optimization of a 20μl PCR system was coupled with the concentration of Mg2+2.0 mmol/L, Taq DNA polymerase activity 0.6U and the annealing temperature 36℃. The DNA fingerprinting of two hybrids was constructed by RAPD marker. Two special bands, A and B, can differentiate Wancao2 and Wancao3 in the fingerprinting.

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