High quality RNA extraction protocol for polyphenolics-rich Cotton tissue

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS Analytical biochemistry Pub Date : 2024-07-08 DOI:10.1016/j.ab.2024.115604

Viralkumar B. Mandaliya , Pritesh Bhatt , Vrinda S. Thaker

引用次数: 0

Abstract

The extraction of high-quality RNA from cotton (Gossypium spp.) is challenging because of the presence of high polyphenolics, polysaccharides, quinones, and other secondary metabolites. A high-throughput RNA extraction protocol is a prerequisite. This Triton-X-100-based RNA extraction method utilizes Polyvinyl pyrrolidone polymer (PVPP) treatment which efficiently removes phenolics, and the application of Lithium chloride (LiCl) has been found that successfully precipitated the high-quality RNA from cotton tissue. Cytoplasmic male sterility (CMS) is a maternally inherited trait associated with specific mitochondrial genome rearrangements or mutations. The suitability of RNA extracted from Cotton CMS lines was assessed. cDNA was synthesized from RNA and assayed for mitochondrial genes (cox3, nad3, nad9) associated with male sterility. This paper discuss the advantages and limitation of this protocol over existing protocol for RNA extraction for polyphenolics-rich plant tissue.

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针对富含多酚的棉花组织的高质量 RNA 提取方案。

从棉花（Gossypium spp.）中提取高质量的 RNA 极具挑战性，因为棉花中含有大量多酚、多糖、醌类和其他次生代谢物。高通量 RNA 提取方案是先决条件。这种基于 Triton-X-100 的 RNA 提取方法采用聚乙烯吡咯烷酮聚合物（PVPP）处理，可有效去除酚类物质，并发现氯化锂（LiCl）的应用可成功沉淀棉花组织中的高质量 RNA。细胞质雄性不育（CMS）是一种与特定线粒体基因组重排或突变有关的母系遗传性状。本文对从棉花 CMS 株系中提取的 RNA 的适用性进行了评估。从 RNA 中合成了 cDNA，并对与雄性不育相关的线粒体基因（cox3、nad3、nad9）进行了检测。本文讨论了该方案与现有的富含多酚植物组织 RNA 提取方案相比的优势和局限性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Analytical biochemistry 生物-分析化学

CiteScore

5.70

自引率

0.00%

发文量

283

审稿时长

44 days

期刊介绍： The journal''s title Analytical Biochemistry: Methods in the Biological Sciences declares its broad scope: methods for the basic biological sciences that include biochemistry, molecular genetics, cell biology, proteomics, immunology, bioinformatics and wherever the frontiers of research take the field. The emphasis is on methods from the strictly analytical to the more preparative that would include novel approaches to protein purification as well as improvements in cell and organ culture. The actual techniques are equally inclusive ranging from aptamers to zymology. The journal has been particularly active in: -Analytical techniques for biological molecules- Aptamer selection and utilization- Biosensors- Chromatography- Cloning, sequencing and mutagenesis- Electrochemical methods- Electrophoresis- Enzyme characterization methods- Immunological approaches- Mass spectrometry of proteins and nucleic acids- Metabolomics- Nano level techniques- Optical spectroscopy in all its forms. The journal is reluctant to include most drug and strictly clinical studies as there are more suitable publication platforms for these types of papers.