一种用于植物基因组DNA提取的低成本高通量方法。

A Low-Cost High-Throughput Method for Plant Genomic DNA Isolation.

作者信息

Gupta Prateek, Salava Hymavathi, Sreelakshmi Yellamaraju, Sharma Rameshwar

机构信息

Repository of Tomato Genomics Resources, Department of Plant Sciences, University of Hyderabad, Hyderabad, India.

出版信息

Methods Mol Biol. 2020;2072:1-7. doi: 10.1007/978-1-4939-9865-4_1.

DOI:10.1007/978-1-4939-9865-4_1

PMID:31541433

Abstract

Many of the functional genomics methods require isolation of genomic DNA from large population of plants. The selection of DNA isolation protocols depends on several factors such as choice of starting material, ease of handling, time and labor required for isolation, the final quantity as well as the quality of genomic DNA. We outline here a high-throughput method of DNA extraction from different plant species including cereal crops. The protocol can be used for extraction of DNA in single tubes as well as for large formats in 96-well plates. The protocol includes steps for eliminating interfering secondary products such as phenolics. This protocol can be applied for high-throughput isolation of DNA for varied applications such as TILLING, mapping, fingerprinting, etc. as a cost-effective protocol compared to commercial kits.

摘要

许多功能基因组学方法都需要从大量植物中分离基因组DNA。DNA分离方案的选择取决于几个因素，如起始材料的选择、操作的简便性、分离所需的时间和劳动力、最终产量以及基因组DNA的质量。我们在此概述一种从包括谷类作物在内的不同植物物种中提取DNA的高通量方法。该方案可用于单管DNA提取，也可用于96孔板的大规模提取。该方案包括消除干扰性次生产物（如酚类物质）的步骤。与商业试剂盒相比，该方案作为一种经济高效的方案，可用于DNA的高通量分离，以用于各种应用，如定向诱导基因组局部突变、作图、指纹识别等。