CAJ | 학술논문

以巨竹叶片提取的基因组DNA为材料，用引物UBC810（序列为GAGAGAGAGAGAGAGAT）研究了PCR反应体系的主要成分、退火温度及循环次数对该种植物ISSR扩增结果的影响。结果表明，20μL的反应体系含40ng模板DNA、0．6μmol·L^-1引物，1．0UTaqDNA聚合酶，2．5mmol·L^-1 Mg^2＋，0．25mmol·L^-1dNTPs，1×Buffer。PCR扩增程序为：94℃预变性5min；94℃变性45s，54．5℃复性30S，70℃延伸90S，循环40次；72℃延伸10min，置4c℃保存。
이거죽협편제취적기인조DNA위재료，용인물UBC810（서렬위GAGAGAGAGAGAGAGAT）연구료PCR반응체계적주요성분、퇴화온도급순배차수대해충식물ISSR확증결과적영향。결과표명，20μL적반응체계함40ng모판DNA、0．6μmol·L^-1인물，1．0UTaqDNA취합매，2．5mmol·L^-1 Mg^2＋，0．25mmol·L^-1dNTPs，1×Buffer。PCR확증정서위：94℃예변성5min；94℃변성45s，54．5℃복성30S，70℃연신90S，순배40차；72℃연신10min，치4c℃보존。
Taking genomie DNA extracted from Gigantochloa levis leaves as the template material, the concentrations of PCR compo- nents, annealing temperature and cycles, whicll affected the ISSR amplification, were optimized with the primer UBC810 （primer sequence ： GAG AGA GAG AGA GAG AT）. The results showed that the 20 μL ISSR reaction system included 40 ng template DNA, 0.6 μmol ·L^-1 primer, 1.0 U Taq DNA polymerase, 2.5 mmol·L^-1 Mg^2＋, 0.25 mmol·L^-1 dNTPs,1×Buffer. The optimal PCR amplification process was 5 minutes at 94 ℃ for predenaturation, followed by 40 cycles ,each with 45 seconds at 94 ℃ for denaturation, 30 seconds at 54.5 ℃ for annealing, 90 seconds at 72℃ for extension,finally extension at 72 ℃ for 10 minutes and holding the samples at 4 ℃.