A high-throughput DNA extraction method for barley seed
A high-throughput DNA extraction method for barley seed
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DOI：
10.1023/a:1022863006134
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发表时间：
2003-01-01
期刊：
影响因子：
1.9
通讯作者：
Tuvesson, S
中科院分区：
文献类型：
--
作者：
von Post, R;von Post, L;Tuvesson, S
描述了一种非破坏性、快速的大麦种子 DNA 提取方法。该方法很简单，包括从种子中钻出样品，添加氢氧化钠，在微波炉中加热并用 Tris-HCl 中和。种子 DNA 提取物可直接用于 PCR，与用于叶子提取物的 PCR 程序相比，在 PCR 程序中添加了额外的循环。该协议是专门针对与大麦黄花叶病毒抗性遗传相关的微卫星标记而开发的，但它也可以应用于大麦育种感兴趣的其他标记。快速种子提取方案使得每天可以处理数千个样品。与叶子样本的长距离转移相比，从种子中提取 DNA 也有利于植物材料的转移。
A non-destructive, quick DNA extraction method for barley seed is described. The method is simple and consists of drilling out a sample from the seed, adding sodium hydroxide, heating in a microwave oven and neutralizing with Tris-HCl. The seed DNA extract can be used directly for PCR with extra cycles added to the PCR programme compared to PCR programmes used for leaf extracts. This protocol was developed in particular for a microsatellite marker genetically linked to barley yellow mosaic virus resistance, but it can be applied to other markers of interest for barley breeding. The quick seed extraction protocol makes it possible to handle thousands of samples per day. Extraction of DNA from seed also facilitates transfer of plant material compared to the long-distance transfer of leaf samples.