入侵植物春飞蓬DNA的提取及ISSR-PCR反应体系的建立

Genomic DNA Extraction Method and ISSR-PCR Amplification Reaction System of Erigeron philadelphicus

以菊科入侵植物的春飞蓬为研究材料,采用CTAB法提取植物总基因组DNA,通过琼脂糖凝胶电泳与紫外分光光度法检测提取的总DNA质量良好,蛋白质、RNA等杂质去除彻底,适合进行ISSR分析;将提取的DNA进行ISSR-PCR扩增,筛选出特异性高的引物,利用单因素和正交试验,建立并优化了入侵植物春飞蓬ISSR-PCR反应体系,20μL优化体系中DNA模板为20 ng、引物为0.2μmol/L、Mg2+为2.5 mmol/L、d NTP为0.2 mmol/L.

This article mainly focused on extraction of DNA with CTAB method from invasive plants of Erigeron philadelphicus. The quality of extraction of DNA was good through agarose gel electrophoresis and ultraviolet spectrophotometry method. Protein and RNA in the DNA extraction were removed completely. The DNA was approved suitable for further ISSR-PCR analysis. High specific primers were screened out. Optimization of the ISSR-PCR reaction system was established by single factor design and orthogonal test design in the experiment. Twenty micro liters ISSR-PCR reaction system includes 20 ng DNA, 0.2 μmol/L primers, 2.5 mmol/L Mg^2＋, 0.2 mmol/L dNTP.