摘要
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。
Using the DNA which extracted from Salix psammophila with TIANGEN as template and the SSR of Salix and Populus as primer, the factors of PCR in reaction system were optimized. The results indicated that, in the optimized PCR reaction system, the most proper combination of 251.d reaction liquid is 40 ng template DNA, 0. 05 U/μl Taq DNA polymerase, 2.5 mmol 10× PCR buffer （ Mg2＋ ）, 0. 25 mmol dNTP, 0. 5μmol primer and enough ddH20 to eke out 251.d reaction liquid. With the help of this system, 30 pairs of Salix psammophila SSR primers were selected. 270 Clones from 9 original areas were amplified by 12 primers, percentage of polymorphic band is 90. 4%. It＇s proves that the SSR of Salix and Populus can be used in Salix psammophila.
出处
《内蒙古农业大学学报（自然科学版）》
CAS
北大核心
2013年第4期59-63,共5页
Journal of Inner Mongolia Agricultural University(Natural Science Edition)
基金
国家自然科学基金(31160167)
教育部高等学校博士点基金(20091515110005)
关键词
北沙柳
SSR—PCR
反应体系优化
引物筛选
Salix psammophila
SSR - PCR
optimization of reaction system
primer screening
