优化的CTAB法提取绿海藻基因组DNA

IF 2.7 3区 生物学 Q2 PLANT SCIENCES
Riyad Hossen, Myles Courtney, Alasdair Sim, Md Abdullah Al Kamran Khan, Heroen Verbruggen, Trevor Bringloe
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引用次数: 0

摘要

海藻正在获得大量的研究兴趣,特别是在基因组应用中,高质量的DNA是先决条件。从这些生物中提取DNA面临着挑战,因为它们的细胞结构不同,生物大分子含量高。现有的方案往往缺乏通用性,导致不同材料和分类群的结果不一致,这突出了对各种绿海藻样品使用的通用方法的需求。方法与结果优化了传统十六烷基三甲基溴化铵(CTAB)提取绿海藻DNA的工艺。我们的方法包括初始样品处理、裂解缓冲液调整和酶孵育改变,在DNA产量和纯度方面优于传统的CTAB和商业试剂盒。值得注意的是,该协议的有效性在各种绿藻材料和保存方法中得到了证明,并在下游应用中进行了测试,结果令人满意。结论优化后的CTAB方案为高质量提取各种绿海藻样品基因组DNA提供了可靠的解决方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

An optimized CTAB method for genomic DNA extraction from green seaweeds (Ulvophyceae)

An optimized CTAB method for genomic DNA extraction from green seaweeds (Ulvophyceae)

Premise

Seaweeds are gaining substantial research interest, particularly for genomic applications, where high-quality DNA is a prerequisite. Extracting DNA from these organisms presents challenges due to high levels of biomacromolecules resulting from their diverse cell structures. Existing protocols often lack versatility, leading to inconsistent outcomes across various materials and taxa, which highlights the need for a universal method for use with a variety of green seaweed samples.

Methods and Results

We optimized the conventional cetyltrimethylammonium bromide (CTAB) protocol for green seaweed DNA extraction. Our method, involving an initial sample treatment, lysis buffer adjustment, and enzyme incubation alterations, outperformed the conventional CTAB and commercial kits in terms of DNA yield and purity. Notably, the protocol's effectiveness was demonstrated across various green algal materials and preservation methods, and was tested with downstream applications with satisfactory results.

Conclusions

Our optimized CTAB protocol offers a reliable solution for high-quality genomic DNA extraction from a wide variety of green seaweed samples.

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来源期刊
CiteScore
7.30
自引率
0.00%
发文量
50
审稿时长
12 weeks
期刊介绍: Applications in Plant Sciences (APPS) is a monthly, peer-reviewed, open access journal promoting the rapid dissemination of newly developed, innovative tools and protocols in all areas of the plant sciences, including genetics, structure, function, development, evolution, systematics, and ecology. Given the rapid progress today in technology and its application in the plant sciences, the goal of APPS is to foster communication within the plant science community to advance scientific research. APPS is a publication of the Botanical Society of America, originating in 2009 as the American Journal of Botany''s online-only section, AJB Primer Notes & Protocols in the Plant Sciences. APPS publishes the following types of articles: (1) Protocol Notes describe new methods and technological advancements; (2) Genomic Resources Articles characterize the development and demonstrate the usefulness of newly developed genomic resources, including transcriptomes; (3) Software Notes detail new software applications; (4) Application Articles illustrate the application of a new protocol, method, or software application within the context of a larger study; (5) Review Articles evaluate available techniques, methods, or protocols; (6) Primer Notes report novel genetic markers with evidence of wide applicability.
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