Genomic DNA Isolation from Maize (Zea mays) Leaves Using a Simple, High-Throughput Protocol

Q1 Agricultural and Biological Sciences

Current protocols in plant biology Pub Date : 2016-05-03 DOI:10.1002/cppb.20000

Kristen A. Leach, Paula C. McSteen, David M. Braun

引用次数: 12

Abstract

A simple, robust, inexpensive, high-throughput method for isolating genomic DNA from maize (Zea mays) leaf tissues is described. The DNA obtained using this extraction protocol is suitable for polymerase chain reaction (PCR) genotyping, which can be employed for the identification of alleles in diverse genetic and breeding approaches, such as marker-assisted selection, genetic fine mapping, and mutant introgression. This method utilizes 96-well plates for the collection of leaf tissue and the subsequent isolation of genomic DNA. The DNA isolation step is performed inexpensively within 3 hr and uses a urea-based extraction buffer that does not require an organic extraction step. Yields of genomic DNA are sufficient to perform ∼25 PCR-genotyping reactions per sample. These qualities, coupled with the protocol being robust and easy for inexperienced users to master, make this method ideal for new researchers. © 2016 by John Wiley & Sons, Inc.

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一种简单、高通量的玉米叶片基因组DNA分离方法

描述了一种简单、可靠、廉价、高通量的方法，用于从玉米(Zea mays)叶片组织中分离基因组DNA。利用该提取方案获得的DNA适合于聚合酶链反应(PCR)基因分型，可用于标记辅助选择、遗传精细作图、突变基因渗入等多种遗传育种方法中的等位基因鉴定。该方法利用96孔板收集叶片组织和随后的基因组DNA分离。DNA分离步骤在3小时内廉价地进行，并使用基于尿素的提取缓冲液，不需要有机提取步骤。每个样品的基因组DNA产量足以进行~ 25次pcr -基因分型反应。这些特性，再加上协议的健壮性和对没有经验的用户易于掌握，使这种方法成为新研究人员的理想选择。©2016 by John Wiley &儿子,Inc。

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来源期刊

Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

自引率

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发文量

期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.