摘要
以广西割手密(S. spontanuem L.)79-9为材料,利用SDS法提取基因组DNA,采用正交试验设计优化建立了一套适用于割手密的SSR-PCR反应体系,即在25μL反应体系中包含10×PCR Buffer(Mg2+free)、模板DNA30ng、Mg2+2.5mmol/L、dNTP0.24μmol/L、引物0.6μmol/L、Taq DNA聚合酶1.5U.运用该体系对其它4份割手密进行了检验,扩增带型清晰,证明该体系稳定可靠.
The genomic DNA of Saccharum spontaneum L. cultivar 79-9 in Guangxi was extracted by SDS, and an SSR-PCR amplification system was set up with the orthogonal experimental design. The system was. 10)4 PCR Buffer (Mg^2+ free), 30 ng DNA template, 2.5 mmol/L Mg^2+, 0.24 μmol/L dNTP, 0. 6 μmol/L primer and 1.5 U Taq DNA polymerase, with a total volume of 25μL. When used to test other S. spontanuem L. populations, this optimized SSR-PCR protocol produced distinct banding patterns, thus indicating its stability and reliability. This system can be used in future analysis of genetic diversity and genetic relationship of S. spontanuern L..
出处
《西南大学学报(自然科学版)》
CAS
CSCD
北大核心
2010年第6期11-16,共6页
Journal of Southwest University(Natural Science Edition)
基金
国家自然科学基金资助项目(30860150)
广西自然科学基金资助项目(Gui Ke Zi 0991009Z
0542026
Gui Ke Qing 0832059)
广西甘蔗研究所基本科研业务专项资助项目(G2009008)