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[目的]对油楠基因组DNA提取、ISSR反应体系优化及引物筛选进行探讨。[方法]运用改良CTAB法对油楠树皮进行基因组DNA提取,利用单因素试验,对油楠ISSR-PCR反应各因素水平进行优化。[结果]运用改良CTAB法对油楠树皮进行基因组DNA提取效果最佳;最优体系:20μl反应体系中,模板用量20 ng,引物浓度0.6μmol/L,d NTPs浓度0.2 mmol/L,Mg2+浓度2.0 mmol/L,Taq酶用量1.5 U,10倍反应缓冲液2μl,dd H2O补至20μl;以该体系为基础进行引物筛选,在100条ISSR随机引物中筛选出13条多态性较高、扩增条带清晰、重复性好的引物。[结论]该研究丰富了油楠遗传学基本资料,为油楠遗传多样性研究奠定基础。
[Objective] The research aimed to explore genomic DNA extraction, ISSR reaction system optimization and primer screening. [Method] The improved CTAB method was used to extract genomic DNA from the bark of C. canola and the factors of ISSR-PCR reaction were optimized by single factor test. [Result] The best CTAB method was used to extract genomic DNA from the bark of C. oleifera. The optimal system was 20 ng of the template, 20 ng of the template, 0.6 μmol / L of the primer, 0.2 mmol / L of dNTPs, Concentration of 2.0 mmol / L, Taq enzyme dosage 1.5 U, 10 times the reaction buffer 2μl, dd H2O make up to 20μl; based on the system primer screening, screening of 100 ISSR random primers 13 high polymorphism , Amplified bands with clear, reproducible primers. [Conclusion] The study enriched the basic information of heliotropia genetics and laid the foundation for the genetic diversity of heliocarpus.