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魔芋属天南星科魔芋属草本植物,其组织富含多糖和多酚等次生物质,这类物质的存在不仅使DNA提取变得困难,还会影响下游的分子生物学操作。采用改进的提取方法,主要包括在2mLEppendorf离心管中液氮研磨,CTAB-SDS裂解细胞,β-巯基乙醇浓度从2%提高到5%和添加PVP抑制多酚氧化,缓冲液去多糖等步骤,成功的提取了魔芋鲜叶的高质量DNA,其A260/A280位于1.80～2.00之间,产率超过470μg/g,ISSR-PCR扩增效果好。该方法具有快速、低廉、微量、稳定等特点,为深入研究魔芋资源遗传多样性、标记辅助育种和种芋纯度的鉴定奠定了基础。
Amorphophallus Amorphophallus Konjac Herb is rich in polysaccharides and polyphenols and other secondary substances, the existence of such substances not only makes DNA extraction more difficult, but also affect the downstream molecular biology operations. The improved extraction method mainly includes the steps of liquid nitrogen grinding in 2mLEppendorf centrifuge tube, cell lysing with CTAB-SDS, increasing the concentration of β-mercaptoethanol from 2% to 5%, adding polyphenol oxidation inhibiting PVP and removing polysaccharides from the buffer, The high-quality DNA of konjac fresh leaves was successfully extracted. The A260 / A280 was located between 1.80 and 2.00 and the yield exceeded 470 μg / g. The ISSR-PCR amplification effect was good. The method has the characteristics of fast, low cost, trace and stability, which lays the foundation for further study on the genetic diversity of konjac resources, marker-assisted breeding and the identification of species purity.