黑核桃叶片基因组DNA提取方法比较研究

Comparison of DNA Extraction Methods from Leaves of Black Walnuts

探索从富含多酚和多糖等次生代谢物质的黑核桃叶片中获得高质量DNA的提取方法。以黑核桃与核桃叶片为材料,采用SDS、CTAB和改良CTAB3种DNA提取方法对黑核桃与核桃基因组DNA提取效果进行比较研究,并通过琼脂糖凝胶电泳与紫外分光光度计检测所提取样品的质量;SDS与CTAB法重复性较差,提取的黑核桃DNA易降解;改良CTAB法可有效抑制酚类物质及细胞内多糖对DNA纯度的影响,对样品提取的DNA也无降解现象,纯度较高,得到的DNAA260/A280值均处于1.80～1.95之间;改良CTAB法是适用于高质量黑核桃DNA的提取方法。

The aim was to obtain the high quality DNA extracted method from leaves of black walnut which contained polysaccharides and polyphenolic compounds.Using leaves of Black walnut and walnut as experimental materials,the genomic DNA extraction of three methods namely SDS,CTAB and modified CTAB were compared.The DNA samples extracted by three methods were tested by agarose gel electrophoresis,ultraviolet spectrophotometer.The results showed that repeatability of the methods of SDS and CTAB were poor,it was difficult to obtain clear bands by agarose gel electrophoresis.Utilizing modified CTAB method,formation of oxidized polyphenolic compounds could be inhibited and most of polysaccharides in the cytoplasmic could be removed efficiently in leaves.A 260 /A 280 was ranged from 1.80-1.95.The modified CTAB method was suitable for extracting high qualified DNA.