高通量植物 DNA 提取在基因组学应用中的研究进展。

High-Throughput DNA Isolation in Vegetable Crops for Genomics Applications.

机构信息

Council for Agricultural Research and Economics - Research Centre for Vegetable and Ornamental Crops (CREA-OF), Pontecagnano, SA, Italy.

CREA Research Centre for Vegetable and Ornamental Crops, Pontecagnano Faiano, Italy.

出版信息

Methods Mol Biol. 2021;2264:47-53. doi: 10.1007/978-1-0716-1201-9_4.

DOI:10.1007/978-1-0716-1201-9_4

PMID:33263902

Abstract

Isolating high-quality DNA is essential for several applications in molecular biology and genomics. Performing whole-genome sequencing in crops and development of reduced representation genomic libraries for genotyping require precise standard on DNA in terms of concentration and purity. For screening large populations it is essential to increase the extraction throughput at affordable costs. In this chapter a homemade protocol is provided that is able to isolate in 96-well plates 198 samples of DNA in a single extraction. The method has been validated in tomato and pepper and can be applied in several vegetable species.

摘要

从分子生物学和基因组学的多个应用来看，提取高质量的 DNA 至关重要。在作物中进行全基因组测序以及开发用于基因分型的简化基因组文库，都需要在浓度和纯度方面对 DNA 有精确的标准。为了对大量群体进行筛选，以经济实惠的成本提高提取通量是很有必要的。本章提供了一个自制的方案，能够在单次提取中从 96 孔板中分离 198 个 DNA 样本。该方法已在番茄和辣椒中得到验证，可应用于多种蔬菜物种。