高质量枣树基因组DNA提取方法的研究

Extraction Ways of High Qualitiful DNA from Z. jujuba Mill.

主要针对枣树组织中多糖严重干扰基因组DNA提取质量的问题,本研究比较了常规CTAB法、TE3D法和改良CTAB法3种方法的处理效果。结果表明:常规CTAB法提取DNA难以去除植物组织中的多糖类杂质;TE3D法提取DNA多糖杂质少,但产率低、易降解、褐化严重;而改良CTAB法提取DNA量大(1000ng/滋l左右),产率高,可达120滋gDNA/g新鲜组织,无明显降解,杂质少,A260/A280比值1.80左右,通过基因组DNA-AFLP指纹图谱分析,完全满足实验要求。并进一步对改良CTAB法的关键步骤作出具体分析讨论。

Based on the question of polyhexose polluting the quality of Z. jujuba Mill. genomic DNA extracted seriously, this study contrasted the extraction effects treateded by three methods of the conventional CTAB, TE3D and improved CTAB entirely. The results showed: The conventional CTAB method was difficult to get rid of polyhexose completely; The TE3D method could remove polyhexose effectively, but extraction rate was very low, DNA browned and degraded seriously; However, the improved CTAB method was an effective way to solve all kind of problem, DNA concentration and extraction rate were high, 1 000ng/?滋l and 120?滋g DNA/g fresh tissue individually, degradation was free nearly, impurity was little, A260/A280 was 1.80 or so. When tested through the DNA-AFLP fingerprinting, experiment requirement was met perfectly. In addition, the key steps and main characters in the improved CTAB method were analyzed in detail.