A Modified CTAB Method for Genomic DNA Extraction from Apple Fruit

生物 基因组DNA DNA DNA提取 萃取(化学) 遗传学 计算生物学 生物技术 基因 色谱法 聚合酶链反应 化学
作者
Jiuying Yan,MA Chang-qing,Chang Bo,Xianguang Fan,Zheng Li,Yang Yazhou,Zhengyang Zhao
出处
期刊:Molecular plant breeding [Sophia Publishing Group, Inc.]
被引量:5
标识
DOI:10.5376/mpb.2018.09.0005
摘要

According to the high content of polysaccharides and polyphenols in apple fruits, three modified CTAB methods were used to extract genomic DNA of ‘Granny Smith’, with young leaves as control. The results showed that the method three was the best. This method integrated multiple purification steps, adding the interferent scavenger before cell lysis, increasing the concentration of β-mercaptoethanol and PVP to inhibite the oxidation of polyphenol, and adding 3 mol/L NaAC of 1/10 volume, which effectively removed the polysaccharides, polyphenols and other secondary metabolites. The results of ultraviolet spectrophotometer and agarose gel electrophoresis showed that DNA was high purity and good integrity. The results of ISSR primers amplification showed that the bands were clear. The results of the digestion of EcoR I restriction enzyme indicated that DNA could be digested by restriction enzyme. Therefore, the method three is the best for genomic DNA extraction. This method provides a reference for the removal of polysaccharides and polyphenols in the process of DNA extraction.

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