CAJ | 학술논문

[目的]筛选经济、稳定性好的水稻PCR反应体系,并检测所选体系在不同的基于PCR反应的分子标记中的通用性.[方法]以CTAB法提取的水稻叶片DNA为模板,应用L_(16)(4~5)正交设计进行PCR反应体系优化.[结果]16个不同处理组合均扩增出了清晰谱带,但扩增效果及PCR产量有差异.最经济、适用的体系:20 μl反应体系,20 ng模板DNA,浓度150 μmol/L dNTP,浓度0.2 μmol/L引物,1.0 U Taq DNA 聚合酶,浓度1.5 mmol/L Mg~(2+),1×Taq buffer,剩余体积用超纯水补齐.[结论]试验确定了水稻PCR优化反应体系,该体系也适用于一些其他基于PCR反应的标记.
[목적]사선경제、은정성호적수도PCR반응체계,병검측소선체계재불동적기우PCR반응적분자표기중적통용성.[방법]이CTAB법제취적수도협편DNA위모판,응용L_(16)(4~5)정교설계진행PCR반응체계우화.[결과]16개불동처리조합균확증출료청석보대,단확증효과급PCR산량유차이.최경제、괄용적체계:20 μl반응체계,20 ng모판DNA,농도150 μmol/L dNTP,농도0.2 μmol/L인물,1.0 U Taq DNA 취합매,농도1.5 mmol/L Mg~(2+),1×Taq buffer,잉여체적용초순수보제.[결론]시험학정료수도PCR우화반응체계,해체계야괄용우일사기타기우PCR반응적표기.
[Objective]The research aimed to screen out the economic and stable PCR system for rice and detect the generality of the selected system in different molecular markers based on PCR. [Method]DNA extracted from rice leaves by CTAB method was used as template to optimize PCR system by using L_(16)(4~5) orthogonal design. [Result]Clear bands were amplified from 16 different combinations, but the amplification effects and yield had difference. The most economic and μmol/L primer, 1.0 U Taq DNA polymerase, 1.5 mmol/L Mg~(2+) , 1×Taq buffer, adding ddH_2O up to terminal volume 20.0 μl. [Conclusion]The experiment confirmed PCR optimized system for rice and the system was slso suitable for other markers based on PCR.