极度濒危植物猪血木RAPD反应体系的优化

采用改进的CTAB法提取中国特有极度濒危植物猪血木(Euryodendron excelsum H.T.Chang) 嫩叶总DNA,并对其进行RAPD分析.分别检测了模板DNA、引物、dNTP、Mg2+和Taq DNA聚合酶用量对反应结果的影响.筛选并建立了适合于猪血木RAPD扩增的反应体系:总体积25 μL,其中10×PCR 缓冲液(500 mmol/L KCl,100 mmol/L Tris-HCl,110% Triton X-100,20 mmol/L MgCl2) 2.5 μL,引物( 1.5 μmol/L) 0.3 μL,dNTPs (10 mmol/L) 0.5 μL,模板2 μL (25 ng),Taq 酶0.2 μL (0.5 U).

Optimal Choice for Reaction System in RAPD Analysis of Euryodendron Excelsum H.T.Chang

The DNA templates of Euryodendron excelsum H.T.Chang extracted from young leaf were amplified by RAPD.Effects of the content of DNA temlates,primers,dNTP,Mg²⁺ and Taq DNA polymerase on experimental results were tested and the optimal reaction system of RAPD for Euryodendron excelsum H.T.Chang was determined as follows:25　ng DNA template,2.0 mmol/L Mg²⁺,0.018　μmol/L primer,250 μmol/L dNTP,0.5 u Taq DNA polymerase in total 25　μL reaction volume.