摘要
以棕榈科植物的DNA为材料,通过对SSR反应体系中模板DNA、dNTPs、引物、Taq聚合酶和MgCl25个因素采用单因素试验法设定5个梯度,并比较不同的浓度、不同的用量对扩增效果的影响,建立最佳反应体系。研究最终确定了20μL PCR反应体系的最佳条件为:模板DNA 2μL,dNTPs 0.2μL,引物1.0μL,Taq聚合酶0.2μL,MgCl22.0μL。该优化体系稳定可靠,适合应用于棕榈科植物SSR分析。
To establish the optimal SSR - PCR reaction system for palm trees, different factors that mostly affected the amplifi- cation of SSR - PCR reaction, including DNA template, dNTPs, primers, TaqDNA and MgC12, were investigated and optimized by setting up 5 gradients with single - fact test method in the experiment. Meanwhile, we also compared the influences of different con centrations and different consumptions on the amplification. The results showed that the optimal SSR - PCR system was DNA template 2 μL, dNTPs 0.2 μL, primers 1.0 μL, TaqDNA 0.2 μL, and MgC12 2.0 μL, which was more stable and suitable for the SSR analysis of palm plants.
出处
《江西农业学报》
CAS
2013年第11期6-10,共5页
Acta Agriculturae Jiangxi
基金
海南省自然科学基金(No.312039)
关键词
棕榈科
SSR
优化
Palm tree
SSR
Optimization