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以TIANGEN试剂盒法提取北沙柳嫩叶的DNA为模板,利用柳属和杨属的SSR引物,建立了北沙柳SSRPCR反应体系,对反应体系中影响聚合酶链式反应(PCR)的因素进行了优化。结果表明,在25μl的PCR反应体系中,DNA模板用量为40ng,TaqDNA聚合酶0.05U/μl,10×PCR buffer(含Mg2+)2.5mM,dNTP 0.25mM,SSR引物0.5μM,加ddH2O补足至25μl时结果最好。利用该体系初步筛选出30对适合北沙柳SSR扩增的引物,并采用优化筛选出的12对引物对9个产地的270个北沙柳无性系进行PCR扩增,多态位点百分率达到90.4%,表明柳属和杨属的SSR引物可以在北沙柳上应用。
The DNA of S. sativa leaves was extracted by TIANGEN kit method. The SSR primers were used to establish SSR PCR reaction system of Salix psammophila and the genus Populus was used as the template. The factors affecting the polymerase chain reaction (PCR) in the reaction system Optimized. The results showed that DNA template dosage was 40 ng, Taq DNA polymerase 0.05 U / μl, 10 × PCR buffer (containing Mg 2+) 2.5 mM, dNTP 0.25 mM and SSR primer 0.5 μM in 25 μl PCR reaction system. When the best results. Thirty pairs of primers suitable for SSR amplification of S. asiatica were initially screened using this system and 270 pairs of P. asarum clones in 9 producing areas were amplified by PCR using 12 pairs of primers optimized, 90.4%, indicating that Salix and Populus SSR primers can be applied in the North Salix.