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相关概念视频

RNA-seq03:21

RNA-seq

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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...
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Ribosome Profiling02:24

Ribosome Profiling

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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...
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相关实验视频

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Probing RNA Structure with Dimethyl Sulfate Mutational Profiling with Sequencing In Vitro and in Cells
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基于质谱的mRNA序列映射通过互补的RNase消化和定制可视化工具.

Emma N Welbourne1, Royce J Copley1, Gareth R Owen1

  • 1School of Chemical, Materials and Biological Engineering, University of Sheffield, Sheffield, UK. m.dickman@sheffield.ac.uk.

The Analyst
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概括

这项研究引入了一种使用部分RNA消化和用于快速mRNA测序的新软件的新方法. 这种方法显著改善了mRNA治疗的序列覆盖和分析准确性.

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科学领域:

  • 生物化学 生物化学
  • 分子生物学分子生物学
  • 生物信息学是一种生物信息学.

背景情况:

  • 使者RNA (mRNA) 技术彻底改变了疫苗开发,大大缩短了时间表.
  • 对mRNA治疗药物的准确表征对于制造和质量控制至关重要.

研究的目的:

  • 开发和验证用于表征mRNA治疗的快速,高吞吐量工作流.
  • 为优化分析和可视化mRNA序列映射创建新型软件.

主要方法:

  • 使用了mRNA的部分RNase T1和RNase U2消化剂.
  • 实现了用于分析的自动化,高吞吐量工作流.
  • 为LC-MS/MS数据处理和可视化开发定制软件.

主要成果:

  • 通过多个RNase T1消化剂达到64%的组合mRNA序列覆盖率,通过组合RNase T1和U2消化剂达到88%.
  • 开发的软件自动化了消化组合,提高了分析速度和准确性.
  • 新的可视化工具,包括螺旋图,改善了序列图分析.

结论:

  • 集成的工作流程和软件能够快速准确地绘制mRNA疗法的序列.
  • 这种方法通过增加覆盖范围和重叠碎片分析,提高了对序列映射的信心.
  • 这种方法促进了对mRNA治疗开发至关重要的高效表征.