Rapid and efficient protocol for genomic DNA extraction from leaf tissues of coconut (Cocos nucifera L.)

Shri Hari Prasad, Lavale Shivaji Ajinath, Deepu Mathew
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引用次数: 1

Abstract

Good quality of the nucleic acid is the primary requisite for genomic research of crop plants. The presence of lipids, polysaccharides, polyphenols and protein molecules hinders downstream processes where genomic DNA has to be used as a template. Coconut leaf being highly fibrous and rich in all the secondary metabolites, isolation of good quality DNA remains a great challenge. Attempts to isolate the coconut DNA following the reported protocols are found not to yield DNA in the expected quality and quantity. A simple and fast approach for isolating the high-quality DNA from polysaccharides and polyphenolic-rich tissues of coconut is being detailed. As measured by its clear color, viscosity, and A260/280 ratio, the isolated DNA was devoid of polysaccharides, polyphenols, RNA, and other significant impurities. In addition to the detailing of the modifications made in the CTAB method, this paper discusses the major step-by-step improvements among the widely-followed DNA isolation protocols.
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快速高效提取椰子叶组织基因组DNA的方法
高质量的核酸是作物基因组研究的首要条件。脂质、多糖、多酚和蛋白质分子的存在阻碍了下游过程,在这些过程中,基因组DNA必须被用作模板。椰子叶纤维含量高,富含所有次生代谢物,因此分离优质DNA仍然是一个巨大的挑战。按照报道的方案分离椰子DNA的尝试被发现不能产生预期质量和数量的DNA。本文详细介绍了一种从椰子多糖和富含多酚的组织中分离高质量DNA的简单快速方法。通过其清晰的颜色、粘度和A260/280比来测量,分离的DNA不含多糖、多酚、RNA和其他重要杂质。除了详细介绍CTAB方法的修改外,本文还讨论了广泛采用的DNA分离协议中主要的逐步改进。
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