CAJ | 학술논문

[目的]寻求高质量枸杞DNA的有效提取方法, 为开展我国枸杞种质资源准确的分子鉴定评价提供试验基础.[方法]针对枸杞组织中多糖严重干扰基因组DNA 提取质量的问题,比较了常规十六烷基三甲基溴化铵(CTAB)法和改良CTAB法的处理效果.[结果]常规CTAB 法提取DNA 难以除去多糖类杂质,溶液极黏稠,致使移液枪吸取困难, 取量不准;改良CTAB 法提取DNA 产率高, 无明显降解, 杂质少, OD_(260 nm)/OD_(280 nm)比值在1.80 左右, 表明DNA 纯度高, 污染少.通过基因组DNA-AFLP 指纹图谱分析, 改良CTAB 法完全满足试验要求.并对改良CTAB 法的关键步骤作了分析讨论.[结论]为枸杞DNA的有效提取提供了方法依据.
[목적]심구고질량구기DNA적유효제취방법, 위개전아국구기충질자원준학적분자감정평개제공시험기출.[방법]침대구기조직중다당엄중간우기인조DNA 제취질량적문제,비교료상규십륙완기삼갑기추화안(CTAB)법화개량CTAB법적처리효과.[결과]상규CTAB 법제취DNA 난이제거다당류잡질,용액겁점주,치사이액창흡취곤난, 취량불준;개량CTAB 법제취DNA 산솔고, 무명현강해, 잡질소, OD_(260 nm)/OD_(280 nm)비치재1.80 좌우, 표명DNA 순도고, 오염소.통과기인조DNA-AFLP 지문도보분석, 개량CTAB 법완전만족시험요구.병대개량CTAB 법적관건보취작료분석토론.[결론]위구기DNA적유효제취제공료방법의거.
[Objective] The basis of the molecular identification and evaluation of Chinese wolfberry(Lycium Linn) resource in China was provided through the research on the good-quality DNA extracted from it. [Method] In order to solve the problem of polyhexose interfering the genomic DNA extracted from Lycium Linn., 2 methods, the conventional CTAB and improved CTAB, were compared. [Results] The results showed that it was difficulty to completely remove the component of polyhexose in the method of conventional CTAB so that there was inaccurate amount in solution-taking because the pipette did not work well under the condition of very sticky liquid. The high-yielding DNA extraction could be realized in the method of modified CTAB, in which there was no significant degradation and less impurity component. The ratio of OD_(260 nm)/OD_(280 nm) was around 1.80, indicating that DNA had high purity and less pollution. Based on the analysis of genomic DNA-AFLP fingerprinting, the improved CTAB method could fully meet the test requirements. As well as a key step in improved CTAB method were analyzed and discussed. [Conclusion] The basis of the effective method of DNA extracted from Chinese wolfberry was provided.