从富含次生代谢物的红树林根组织中高效分离RNA的改进方法。

IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS BioTechniques Pub Date : 2023-06-01 DOI:10.2144/btn-2022-0078
Ashifa Nizam, Haritha Kalath, Ajay Kumar
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引用次数: 0

摘要

红树林中的次生代谢物经常干扰RNA的提取,导致浓度和质量差,不适合下游应用。由于现有的方法只能从candelia canddel (L.)的根组织中提取低质量的RNA。山药与大根霉。为提高RNA的质量和产率,提出了一种优化的方法。与其他三种方法相比,该优化方案对两种物种都具有更高的RNA产量和纯度。A260/280和A260/230的吸光度比≥1.9,RNA完整性值为7.5 ~ 9.6。结果表明,该方法可以有效地从红树林根系中提取高质量的RNA,适用于cDNA合成、实时定量PCR和下一代测序等下游实验。
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A modified method for efficient RNA isolation from mangrove root tissues rich in secondary metabolites.

Secondary metabolites in mangroves often interfere with RNA extraction yielding poor concentration and quality, which is unsuitable for downstream applications. As existing protocols yielded low-quality RNA from root tissues of Kandelia candel (L.) Druce and Rhizophora mucronata Lam., an optimized method was developed for improving the quality and yield of RNA. Compared with three other methods, this optimized protocol gave better RNA yield and purity for both species. The absorbance ratios were ≥1.9 for A260/280 and A260/230, while RNA integrity number values ranged from 7.5 to 9.6. Results show that our modified method is efficient in obtaining high-quality RNA from mangrove roots and is suitable for downstream experiments such as cDNA synthesis, real-time quantitative PCR and next-generation sequencing.

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来源期刊
BioTechniques
BioTechniques 工程技术-生化研究方法
CiteScore
4.40
自引率
0.00%
发文量
68
审稿时长
3.3 months
期刊介绍: BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.
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