胡卢巴RAPD反应体系优化及应用

Applying and optimization of RAPD reaction system in Trigonella foenum-graecum

以胡卢巴DNA为模板,采用随机引物,优化胡卢巴的RAPD反应体系和反应条件.结果表明,在20μL反应体系中,5种主要因素Taq DNA聚合酶、引物、dNTPs、模板DNA的最适浓度和退火温度分别是1μmol/min,15μmol/L,0.30 mmol/L,20 ng和34℃.在优化的RAPD反应体系条件下,构建22个胡卢巴种质的RAPD指纹图谱,为胡卢巴遗传多样性检测和其他方面的应用提供分子标记手段.

DNA from Trigonella foenum-graecum was template, the optimal reaction of RAPD in Trigonella foenum-graecurn was studied with RAPD primer. The results showed that the optimum concentration of five important components i. e. Taq DNA polymerase, primer, dNTPs , template DNA and annealing temperature in 20 μL reaction system were 1 μmol/min, 15 μmol/L, 0. 30 mmol/L, 20 ng and 34 ℃ respectively. The RAPD finger-printing of 22 Trigonella foenum-grdecum germplasms were established by using optimum system, which offered method of molecular marker to genetic diversity measuring and other application in Trigonella foenum-graecum.