CAJ | 학술논문

以大花君子兰为材料研究了PCR反应体系的主要成分及退火温度对君子兰ISSR扩增结果的影响,并对影响PCR反应体系的主要成分及退火温度进行筛选和优化,确立了适合君子兰ISSR-PCR分析的最佳反应体系:在20 μL的反应体系中,Mg~(2+)为2.0 mmol/L,模板DNA用量为50 ng/μL,Taq酶0.4 U,dNTPs浓度为0.15 mmol/L,引物浓度为0.4 mmol/L.筛选出了13 个适宜君子兰遗传分析的ISSR引物,确定最适宜退火温度为52～56℃.
이대화군자란위재료연구료PCR반응체계적주요성분급퇴화온도대군자란ISSR확증결과적영향,병대영향PCR반응체계적주요성분급퇴화온도진행사선화우화,학립료괄합군자란ISSR-PCR분석적최가반응체계:재20 μL적반응체계중,Mg~(2+)위2.0 mmol/L,모판DNA용량위50 ng/μL,Taq매0.4 U,dNTPs농도위0.15 mmol/L,인물농도위0.4 mmol/L.사선출료13 개괄의군자란유전분석적ISSR인물,학정최괄의퇴화온도위52～56℃.
The factors affecting the ISSR (inter-simple sequence repeat) result of Clivia (Clivia miniata) were studied.By adjusting Mg~(2+),dNTPs,primer,Taq DNA polymerase and annealing temperature were optimized.The optimal ISSR-PCR reaction conditions for Clivia miniata were determined as follows,in 20 μL PCR reaction system,2.0 mmol/L Mg~(2+),50 ng/μL template DNA,0.4 U Taq DNA polymerase,0.15 mmol/L dNTPs,0.4 μmol/L primer.Thirteen ISSR primers that are suitable for Clivia were screened,and the optimal annealing temperature for the ISSR reaction was 52-56℃.