A high throughput DNA extraction method with high yield and quality.
A high throughput DNA extraction method with high yield and quality.
复制标题
DOI：
10.1186/1746-4811-8-26
复制
发表时间：
2012-07-28
期刊：
影响因子：
5.1
通讯作者：
Chen J
中科院分区：
文献类型：
--
作者：
Xin Z;Chen J
从大量植物样品中制备大量、高质量的基因组 DNA 是大多数遗传和基因组分析的主要瓶颈，例如遗传作图、TILLING（基因组中靶向诱导的局部损伤）和直接从剪切的基因组 DNA 进行下一代测序。有多种 DNA 制备方法和商业试剂盒可供选择。然而，它们要么产量低、产量低，要么成本高。在这里，我们描述了一种从高粱 [Sorghum bicolor (L.) Moench] 叶和干种子中高通量分离基因组 DNA 的方法，具有高产量、高质量和可承受的成本。我们将高产量 CTAB 提取方法与基于 MagAttract 试剂盒的改进净化程序相结合，开发了一种高通量 DNA 分离方法。该方法从冻干高粱叶和干种子中获得了大量、高质量的 DNA。 DNA 产量提高了近 30 倍，MagAttract 磁珠的消耗量减少了 4 倍。该方法也可用于其他植物物种，包括棉叶和松针。开发并验证了从高粱叶和种子中提取 DNA 的高通量系统。该方法的主要优点是成本低、产量高、质量高、通量大。一个人可以在一个工作日内处理两个 96 孔板，每个磁珠样本的成本为 0.10 美元，外加其他方法也需要的其他消耗品。
Preparation of large quantity and high quality genomic DNA from a large number of plant samples is a major bottleneck for most genetic and genomic analyses, such as, genetic mapping, TILLING (Targeting Induced Local Lesion IN Genome), and next-generation sequencing directly from sheared genomic DNA. A variety of DNA preparation methods and commercial kits are available. However, they are either low throughput, low yield, or costly. Here, we describe a method for high throughput genomic DNA isolation from sorghum [Sorghum bicolor (L.) Moench] leaves and dry seeds with high yield, high quality, and affordable cost. We developed a high throughput DNA isolation method by combining a high yield CTAB extraction method with an improved cleanup procedure based on MagAttract kit. The method yielded large quantity and high quality DNA from both lyophilized sorghum leaves and dry seeds. The DNA yield was improved by nearly 30 fold with 4 times less consumption of MagAttract beads. The method can also be used in other plant species, including cotton leaves and pine needles. A high throughput system for DNA extraction from sorghum leaves and seeds was developed and validated. The main advantages of the method are low cost, high yield, high quality, and high throughput. One person can process two 96-well plates in a working day at a cost of $0.10 per sample of magnetic beads plus other consumables that other methods will also need.
登录
查看更多内容
影响因子：
2.3
作者：
Flagel, L;Christensen, JR;Matthews, PD
通讯作者：
Matthews, PD
影响因子：
7.4
作者：
Henikoff, S;Till, BJ;Comai, L
通讯作者：
Comai, L
影响因子：
2.1
作者：
Dilworth, E;Frey, JE
通讯作者：
Frey, JE
影响因子：
2
作者：
Abd-Elsalam, K. A.;Asran-Amal, A.;El-Samawaty, A. M. A.
通讯作者：
El-Samawaty, A. M. A.
影响因子：
2.9
作者：
Michiels, A;Van den Ende, W;Van Laere, A
通讯作者：
Van Laere, A