割手密(S. spontanuem L.)SSR-PCR反应体系的优化

Optimization of SSR-PCR System for S. spontanuem L.

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..