CAJ | 학술논문

为建立思茅松大配子体ISSR分子标记的反应体系,以思茅松种子样本作为供试材料,比较不同思茅松胚乳DNA提取方法的效果,通过紫外分光度计、DNA琼脂糖电泳和PCR扩增分别予以检测并比较。结果表明SDS法优于CTAB法。同时,通过单因子分析法对ISSR-PCR反应体系中的模板DNA用量,Taq DNA聚合酶用量,Mg2＋浓度,dNTP浓度、引物浓度等主要影响因子进行优化,确立适用于思茅松的ISSR分子标记反应体系。对该体系进行稳定性检验,结果表明该反应体系适合思茅松ISSR-PCR扩增。
위건립사모송대배자체ISSR분자표기적반응체계,이사모송충자양본작위공시재료,비교불동사모송배유DNA제취방법적효과,통과자외분광도계、DNA경지당전영화PCR확증분별여이검측병비교。결과표명SDS법우우CTAB법。동시,통과단인자분석법대ISSR-PCR반응체계중적모판DNA용량,Taq DNA취합매용량,Mg2＋농도,dNTP농도、인물농도등주요영향인자진행우화,학립괄용우사모송적ISSR분자표기반응체계。대해체계진행은정성검험,결과표명해반응체계괄합사모송ISSR-PCR확증。
Different methods of DNA extraction were compared by using the ultraviolet spectrophotometer,DNA agarose electrophoresis and PCR amplification,in order to establish the ISSR reaction system of the macrogametocytes of Pinus kesiya var.langbianensis,in which the seeds were used as materials.The results showed that the method of SDS was better than CTAB,and meanwhile,the ISSR reaction system was established by the analysis of the main factors such as the usage of template DNA and Taq DNA polymerase,the concentration of Mg2＋,dNTP and primer.The stability of the ISSR reaction system was tested and the result showed that this reaction system was suited to the ISSR-PCR amplification of Pinus kesiya var.langbianensis.