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RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...

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Updated: Jul 9, 2026

Purifying the Impure: Sequencing Metagenomes and Metatranscriptomes from Complex Animal-associated Samples

Purifying the Impure: Sequencing Metagenomes and Metatranscriptomes from Complex Animal-associated Samples

Published on: December 22, 2014

优化RNA提取用于米散土土样本的转录基因组测序.

Sumitra Sivaprakasam¹, Tan Yee Fan¹, Yukgehnaish Kumarasan¹

¹Centre of Excellence for Omics-Driven Computational Biodiscovery (COMBio), AIMST University, Bedong, Kedah, 08100, Malaysia.

BMC research notes

|July 5, 2025

概括

使用聚乙烯糖醇 (PEG) 沉优化从土中提取RNA,显著提高了RNA产量和质量. 这种增强的方法对于植物生长研究中准确的微生物组分析至关重要.

关键词:

大量的土壤土壤.转基因组组序列的测序.帕迪·帕迪 (Paddy Paddy) 是一个提取RNA 提取RNA的方法

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Last Updated: Jul 9, 2026

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科学领域:

微生物学微生物学
分子生物学分子生物学
植物科学植物科学

背景情况:

高质量的RNA提取对于研究米植物中的微生物群分子动力学至关重要.
米土的湿物质和粘土含量阻碍了RNA提取,导致吸附和低产量.
现有的RNA提取方法需要针对具有挑战性的土壤环境进行优化.

研究的目的:

改进现有的RNA提取协议,用于马来西亚大米田的土壤样本.
为了优化RNA提取高颜色和具有挑战性的米土壤样本.
为了评估优化的RNA适合于转基因组序列的适用性.

主要方法:

对多个已发表的RNA提取协议的评估.
使用聚乙烯糖醇 (PEG) 沉 (20%和30%) 优化选定的协议 (Peng等人,2018年,方法B3).
评估RNA产量,纯度 (A260/A280) 和完整性 (RIN),然后进行元转录组测序.

主要成果:

使用20%的PEG沉的优化方案有效地去除了色素,产生了无色素RNA (>100 ng/μl,完整性>7,A260/A280的2.02±0.02).
转录基因组测序和分析 (Trinity,BUSCO,Kraken2) 证实了优越的RNA质量.
20%的PEG方法显示了更高的细菌读数分配,这表明下游应用的有效性.

结论:

使用20%的PEG沉的优化RNA提取协议对米土壤非常有效.
这种方法显著提高了RNA质量和产量,克服了土壤污染物带来的挑战.
提取的RNA适用于强大的微生物转录组分析和在米生态系统中的微生物组研究.