相关实验视频
Updated: Jul 11, 2025
12:01
3' End Sequencing Library Preparation with A-seq2
Published on: October 10, 2017
10.6K
在基底分辨率下对mRNA pseudouridine的全转录组定量测序进行BID-seq
Li-Sheng Zhang1,2,3,4, Chang Ye1,2, Cheng-Wei Ju2,5
1Department of Chemistry, The University of Chicago, Chicago, IL, USA.
Nature protocols
|November 15, 2023
概括
在mRNA中伪尤里丁 (Ψ) 映射现在是定量性的. 我们优化的双硫酸盐诱导删除测序 (BID-seq) 方法快速识别哺乳动物mRNA中的数千个 Ψ位点.
科学领域:
- 分子生物学分子生物学
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
- 基因组RNA的修改 基因组RNA的修改
背景情况:
- 伪尤里丁 (Ψ) 是一种常见的RNA修饰,在各种RNA类型中发现,包括mRNA.
- 哺乳动物mRNA Ψ的功能研究受到缺乏精确的定量检测方法的限制.
- 现有的方法缺乏为全面的 Ψ 映射所需的基对分辨率.
研究的目的:
- 提出一个优化的二硫酸盐诱导删除序列 (BID-seq) 协议,用于对细胞mRNA中的 Ψ分布进行定量映射.
- 为了实现高分辨率的mRNA伪氨基基化转录组范围的分析.
- 为 Ψ 站点检测和量化提供一个用户友好的数据分析管道.
主要方法:
- 开发了一种优化的BID-seq协议,简化了图书馆准备和数据分析.
- 确保了 Ψ 位点的高删除比率,同时最大限度地减少了未经修改的尿液的背景删除.
- 利用下一代测序 (NGS) 进行全转录组 Ψ 分析.
主要成果:
- 优化的BID-seq协议具有高度可重现性和灵敏性,只需要10 ng的多A+RNA.
- 在小鼠胚胎干细胞中成功确定了8407个 Ψ 位点.
- 相关的BID-pipe软件有助于 Ψ 位点检测和静脉测量量量.
结论:
- 优化的BID-seq协议提供了一种强大,快速和定量方法,用于绘制mRNA伪化.
- 这一工作流显著推进了对mRNA Ψ修饰及其在哺乳动物中的功能作用的研究.
- 协议和分析管道可供具有基本分子生物学和生物信息学技能的研究人员访问.
相关概念视频
03:21RNA-seq
10.0K
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...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
10.0K
02:24Ribosome Profiling
3.5K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.5K