Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan, China.
J Genet Genomics. 2013 Jan 20;40(1):45-54. doi: 10.1016/j.jgg.2012.10.001. Epub 2012 Nov 10.
DNA is one of the most basic and essential genetic materials in the field of molecular biology. To date, isolation of sufficient and good-quality DNA is still a challenge for many plant species, though various DNA extraction methods have been published. In the present paper, a recycling DNA extraction method was proposed. The key step of this method was that a single plant tissue sample was recycled for DNA extraction for up to four times, and correspondingly four DNA precipitations (termed as the 1st, 2nd, 3rd and 4th DNA sample, respectively) were conducted. This recycling step was integrated into the conventional CTAB DNA extraction method to establish a recycling CTAB method. This modified CTAB method was tested in eight plant species, wheat, sorghum, barley, corn, rice, Brachypodium distachyon, Miscanthus sinensis and tung tree. The results showed that high-yield and good-quality DNA samples could be obtained by using this new method in all the eight plant species. The DNA samples were good templates for PCR amplification of both ISSR and SSR markers. The recycling method can be used in multiple plant species and can be integrated with multiple conventional DNA isolation methods, and thus is an effective and universal DNA isolation method.
DNA 是分子生物学中最基本和最重要的遗传物质之一。迄今为止，尽管已经发表了各种 DNA 提取方法，但对于许多植物物种来说，仍然难以获得足够数量和高质量的 DNA。本文提出了一种回收 DNA 提取方法。该方法的关键步骤是，将单个植物组织样本回收用于 DNA 提取，最多可进行四次，相应地进行四次 DNA 沉淀（分别命名为第 1 次、第 2 次、第 3 次和第 4 次 DNA 样品）。将该回收步骤集成到常规 CTAB DNA 提取方法中，建立了回收 CTAB 方法。该改良的 CTAB 方法在 8 种植物物种（小麦、高粱、大麦、玉米、水稻、短柄草、芒草和油桐）中进行了测试。结果表明，该新方法可在所有 8 种植物物种中获得高产和高质量的 DNA 样品。DNA 样品是 ISSR 和 SSR 标记 PCR 扩增的良好模板。该回收方法可用于多种植物物种，并可与多种常规 DNA 分离方法集成，因此是一种有效且通用的 DNA 分离方法。
