梨属植物RAPD反应体系的建立与优化

Establishment and optimization of RAPD technique in Pyrus L.

以鸭梨、杜梨为材料,对梨属植物DNA的提取及RAPD反应体系进行了系统优化,建立了梨属植物最佳反应体系及参数。结果表明,适当提高抗氧化、抗酚类物质的浓度(2%β-巯基乙醇,2%PVP-40,20 mmol/LNa2S2O5),提取的基因组DNA纯度较高;反应体系中含有Mg2+2.0 mmol/L,dNTPs浓度200μmol/L,引物0.4μmol/L,模板DNA 1.6 ng/μL,Taq DNA聚合酶0.06 U/μL,可成功用于梨属植物RAPD扩增。说明改良的CTAB法能成功用于梨属植物DNA提取,建立的最佳反应体系可用于梨属植物RAPD扩增。

Tested with Yali pear and Betulaefolia,the paper studied the extract of DNA of Pyrus and the optimized RAPD reaction system. The result showed as follows .by raising the concentrations of antioxidative components and antiphenolic compounds （2% β-ME,2% PVP-40,20 mmol/L Na2S2O5）,the purity of the extracted genomic DNA was higher. It＇s concluded that the optimum reaction system that contained Mg^2＋ 2.0 mmol/L, dNTPs 200 μmol/L, primer 0.4 μmol/L, 1.6 ng/μL template DNA, 0. 06 U/μL, Taq DNA polymerase would be applied in the amplification of RAPD. It explained that reformed CTAB method could extract the genomic DNA of Pyrus successfully,and the optimum RAPD reaction system could amplificate RAPD in Pyrus L..