CAJ | 학술논문

为研究文冠果种质遗传多样性 ,以新疆奇台文冠果自然栽培居群的48个单株为材料 ,采用改良CTAB法提取文冠果基因组DNA ,并对其RAPD和ISSR反应体系进行了优化.研究结果表明 :文冠果的RAPD最适反应体系为 :PCR扩增总体积为20 μL ,包含30 ng 的模板 DNA ,10 × PCR buffer 2 μL ,2 .0 mmol·L-1 Mg2+ ,0 .1 mmol·L-1 dNTP和 Taq DNA聚合酶1U ;ISSR最适反应体系为 :PCR扩增总体积为20 μL ,包括30 ng的模板DNA ,10 × PCR buffer 2 μL ,2 .5 mmol·L-1 Mg2+ ,0 .2 mmol·L-1 dNTP和 Taq DNA聚合酶1U.在最适反应体系下 ,RAPD和ISSR分析具有良好的稳定性和可重复性.
위연구문관과충질유전다양성 ,이신강기태문관과자연재배거군적48개단주위재료 ,채용개량CTAB법제취문관과기인조DNA ,병대기RAPD화ISSR반응체계진행료우화.연구결과표명 :문관과적RAPD최괄반응체계위 :PCR확증총체적위20 μL ,포함30 ng 적모판 DNA ,10 × PCR buffer 2 μL ,2 .0 mmol·L-1 Mg2+ ,0 .1 mmol·L-1 dNTP화 Taq DNA취합매1U ;ISSR최괄반응체계위 :PCR확증총체적위20 μL ,포괄30 ng적모판DNA ,10 × PCR buffer 2 μL ,2 .5 mmol·L-1 Mg2+ ,0 .2 mmol·L-1 dNTP화 Taq DNA취합매1U.재최괄반응체계하 ,RAPD화ISSR분석구유량호적은정성화가중복성.
In order to study the genetic diversity of X anthoceras Sorbifolia germplasms ,taking 48 individuals as materials growing in natural cultivated population in the Qitai county of Xinjiang germpalsm nursery .Leaves DNA of each sample were extracted using the modified CTAB method ,RAPD and ISSR reaction systems were optimized .The results showed that the optimal RAPD reaction system was in the total volume of 20 μL for PCR amplification ,containing 30 ng template DNA ,2μL 10 × PCR buffer ,2 .0 mmol·L-1 Mg2+ ,0 .1 mmol·L-1 dNTPs ,1U Taq enzyme .The optimal ISSR analysis reaction system was in the total volume of 20 μL for PCR amplification ,containing 30 ng template DNA ,2 μL 10 × PCR buffer ,2 .5 mmol·L-1 Mg2+ ,0 .2 mmol·L-1 dNTPs ,1U Taq enzyme .In these optimum reaction systems ,RAPD and ISSR analysis had stability and repeat-ability very well .