枸杞DNA的提取方法研究

Research on the Extraction Method of High Quality DNA from Chinese Wolfberry

[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,OD260 nm/OD280 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。

[ 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 inaccu- rate 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 0D260nm/OD280 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.