摘要
利用单因素分析法对影响菰SRAP-PCR扩增效果的Mg2+、dNTPs、Taq DNA聚合酶、DNA模板和引物浓度5种因素进行了优化。结果表明:建立了适合菰基因组的SRAP-PCR的体系:1μL 10×Buffer、0.5 U Taq DNA聚合酶、2 mmol/L MgCl2、50 ng模板DNA、0.20 mmol/L dNTPs、正反向引物的浓度各为0.7μmol/L,共10μL。选取5对引物,利用该体系对72份菰样本进行PCR扩增,共获得132条清晰的谱带,其中91条具有多态性,比率为68.9%。菰SRAP-PCR反应体系的优化和建立将为利用该标记进行种质遗传多样性分析和连锁图谱构建等研究提供技术支持。
We established an advanced SRAP-PCR reaction system for Zizania latifolia using a single factor experiment with five impact factors, including DNA template, Mg2＋, dNTPs mixture, Taq DNA polymerase and primer. The 10 p.L reaction mixture contained 50 ng of genomic DNA template, 1 μL of 10×Buffer, 2 mmol/L of MgC12, 0.20 mmol/L of dNTP, 0.5 U of Taq DNA polymerase, and 0.7 μmol/L of primers. To test the stability of the optimized SRAP-PCR system, the PCR was further amplified in Z. latifblia with 72 individuals using 5 primer pairs. A total of 132 clear bands were obtained throughout all materials, of which 91 （68.9%） bands were polymorphic. Therefore, the established SRAP reaction system for Z. latifolia was reliable. The results provided mature technical support for evaluating genetic diversity and constructing genetic linkage maps of Z. latifolia.
出处
《热带作物学报》
CSCD
北大核心
2014年第2期299-306,共8页
Chinese Journal of Tropical Crops
基金
"安徽省农业综合开发项目<蔬菜新品种引进与示范>"(No.11003258)
关键词
菰
SRAP
PCR反应体系
优化
Zizania latifolia
SRAP
PCR reaction system
Optimization
