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微生物组的单个微生物RNA测序使用smRandom-Seq
Ziye Xu1,2, Yuting Wang3, Wenjie Cai1
1Department of Laboratory Medicine of The First Affiliated Hospital and Liangzhu Laboratory, Zhejiang University School of Medicine, Hangzhou, China.
Nature protocols
|May 22, 2025
概括
我们开发了单个微生物RNA测序 (smRandom-seq) 来揭示细菌的多样性. 这种高通量方法分析了个别的微生物细胞,揭示了传统方法错过的异质性.
科学领域:
- 微生物学 微生物学
- 基因组学就是基因组学.
- 分子生物学分子生物学
背景情况:
- 细菌在各种环境中表现出显著的多样性.
- 传统的转录学方法平均基因表达,掩盖个体微生物的行为和异质性.
- 了解微生物异质性对于微生物组研究和传染病等领域至关重要.
研究的目的:
- 引入一种基于滴滴的新型,高通量单微生物RNA测序 (smRandom-seq) 方法.
- 为微生物样本分析实施smRandom-seq提供详细的协议.
- 为了能够在单细胞水平上深入研究微生物异质性.
主要方法:
- 开发了一种基于滴滴的高通量单微生物RNA测序 (smRandom-seq) 协议.
- 程序包括微生物样本预处理,现场预索引cDNA合成,现场聚dA尾巴,滴滴条形编码,核糖体RNA枯竭和图书馆准备.
- 整个工作流程,从样本处理到图书馆建设,大约在2天内完成.
主要成果:
- smRandom-seq实现了高度物种特异和敏感的基因检测.
- 该方法显示了增强的RNA覆盖范围,降低了双倍率,并最大限度地减少了核糖体RNA污染.
- 成功应用于实验室培养和复杂微生物群体样本.
结论:
- smRandom-seq提供了一个强大的工具,用于构建单个微生物的转录组图集.
- 这种方法有助于在不同环境中对微生物异质性的详细分析.
- 它对细菌耐药性,微生物组复杂性和宿主微生物相互作用的研究具有重大潜力.
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