Polymerase chain reaction has found wide applications in modern research involving transformations and other genomic studies. For reproducible PCR results, however, the quantity and quality of template DNA is of considerable importance. A simple and efficient plant DNA extraction procedure for isolation of high-quality DNA from plant tissues is presented here. It requires maceration of plant tissue of about 1.0 cm² (e.g. of a leaf blade) in DNA extraction buffer (100 mM Tris-HCl, 100 mM EDTA, 250 mM NaCl) using 1.5-mL microfuge tubes, followed by cell lysis with 20% SDS, and DNA extraction with phenol: chloroform: iso-amyl alcohol (25:24:1). Hydrated ether is then used to remove polysaccharides and other contaminants from the DNA preparation. Average DNA yield is 20–30 μg cm⁻² for fresh tissues, and ratio of absorbance at 260 nm to absorbance at 280 nm is 1.5–1.8. The DNA is quite suitable for PCR using microsatellites, RAPD and specific markers for recombinant selection. Amplifications have been obtained for these markers by using template DNA extracted from fresh as well as frozen leaf tissues of various plants, including barley, oat, potato and tomato. DNA stored for more than 2 years has been successfully amplified with microsatellite markers, which shows suitability of this method after long-term storage of DNA. Besides, the ease of use and cost-effectiveness make the procedure attractive.

中文翻译：

---

用于 PCR 的高质量植物 DNA 提取：一种简单的方法。

聚合酶链反应在涉及转化和其他基因组研究的现代研究中得到了广泛的应用。然而，对于可重复的 PCR 结果，模板 DNA 的数量和质量非常重要。这里介绍了一种从植物组织中分离高质量 DNA 的简单有效的植物 DNA 提取程序。它需要使用 1.5 mL 离心管在 DNA 提取缓冲液（100 mM Tris-HCl、100 mM EDTA、250 mM NaCl）中浸渍约 1.0 cm ²（例如叶片）的植物组织，然后用 20 % SDS，并用苯酚提取 DNA：氯仿：异戊醇 (25:24:1)。然后使用水合醚从 DNA 制备物中去除多糖和其他污染物。平均 DNA 产量为 20–30 μg cm ^-2对于新鲜组织，260 nm 处的吸光度与 280 nm 处的吸光度之比为 1.5–1.8。DNA 非常适合使用微卫星、RAPD 和用于重组选择的特定标记进行 PCR。已通过使用从各种植物（包括大麦、燕麦、马铃薯和番茄）的新鲜和冷冻叶组织中提取的模板 DNA 获得了这些标记的扩增。使用微卫星标记成功扩增了保存2年以上的DNA，表明该方法在DNA长期保存后的适用性。此外，易用性和成本效益使该程序具有吸引力。