Optimized method for dna extraction and PCR amplification in aroeira tree

IF 0.6 4区农林科学 Q3 AGRICULTURE, MULTIDISCIPLINARY

Bioscience Journal Pub Date : 2023-03-03 DOI:10.14393/bj-v39n0a2023-62577

Jailson do Nascimento Silva, Amanda Camila dos Santos Linhares, D. A. Palmieri, Marcones Ferreira Costa, G. H. Sá, M. Gomes, M. F. Santos, L. Feitoza, Sérgio Emílio Dos Santos Valente

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引用次数: 1

Abstract

Molecular markers are important tools in the characterization of plant genetic diversity and can provide support for conservation strategies for endangered populations. The different molecular techniques involve the evaluation of many individuals; therefore, it is crucial to have fast, efficient, and inexpensive methods for DNA extraction. Given the importance of the Aroeira (Myracrodruon urundeuva Fr. All.) it is pertinent to optimize a protocol that allows the obtainment of intact and pure DNA, aiming to assist conservation strategies for this species that is threatened with extinction. Thus, this study aimed to compare five DNA extraction methods: Dellaporta et al. (1983), Doyle and Doyle (1987) modified, Ferreira and Grattapaglia (1995), Romano and Brasileiro (2015), and Khanuja et al. (1999) and optimize the most efficient protocol for M. urundeuva. The modified DNA extraction protocol proposed by Doyle and Doyle (1987), using 100 mg of leaf tissue and 6 µl of β-mercaptoethanol was the protocol that presented the sharpest bands after DNA electrophoresis and after the reactions of amplification employing Polymerase Chain Reaction (PCR). Therefore, it is suggested to use the protocol described by Doyle and Doyle (1987) modified for the extraction of DNA from young M. urundeuva leaves to carry out techniques involving molecular markers.

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牛头树dna提取及PCR扩增方法优化

分子标记是鉴定植物遗传多样性的重要工具，可以为濒危种群的保护策略提供支持。不同的分子技术涉及对许多个体的评估;因此，寻找快速、高效、廉价的DNA提取方法至关重要。考虑到Aroeira (Myracrodruon urundeuva Fr. All.)的重要性，优化一种能够获得完整和纯DNA的方案是相关的，旨在帮助保护这一濒临灭绝的物种。因此，本研究旨在比较五种DNA提取方法:Dellaporta等人(1983)、Doyle和Doyle (1987) modified、Ferreira和Grattapaglia(1995)、Romano和Brasileiro(2015)和Khanuja等人(1999)，并优化M. urundeuva最有效的提取方案。Doyle和Doyle(1987)提出的改良DNA提取方案，使用100 mg叶组织和6µl β-巯基乙醇，是DNA电泳和PCR扩增反应后条带最清晰的方案。因此，建议采用Doyle和Doyle(1987)所描述的修改后的提取野檀幼叶DNA的方案，进行涉及分子标记的技术。

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来源期刊

Bioscience Journal Agricultural and Biological Sciences-General Agricultural and Biological Sciences

CiteScore

1.00

自引率

0.00%

发文量

审稿时长

48 weeks

期刊介绍： The Bioscience Journal is an interdisciplinary electronic journal that publishes scientific articles in the areas of Agricultural Sciences, Biological Sciences and Health Sciences. Its mission is to disseminate new knowledge while contributing to the development of science in the country and in the world. The journal is published in a continuous flow, in English. The opinions and concepts expressed in the published articles are the sole responsibility of their authors.