降级组序列化 降级组序列化
Shih-Shun Lin1, Yihua Chen2, Mei-Yeh Jade Lu3
1Institute of Biotechnology, National Taiwan University, Taipei, Taiwan. linss01@ntu.edu.tw.
Methods in molecular biology (Clifton, N.J.)
|May 16, 2025
概括
使用cDNA末端 (5'-RACE) 的5'-快速放大和下一代测序 (NGS) 的降解组测序,揭示了RNA降解模式. 这种方法可以识别微RNA的点,并有助于进行RNA处理研究,特别是在非模型生物中.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 生物技术是生物技术.
背景情况:
- RNA降解对基因调节至关重要.
- 了解RNA处理机制对于生物研究至关重要.
- 降级组测序为研究RNA动态提供了一个强大的工具.
研究的目的:
- 为降解组图书馆构建提供一个优化的协议.
- 通过下一代测序 (NGS) 实现降解组cDNA片段的测序.
- 为了方便识别microRNA目标和研究RNA处理.
主要方法:
- 适应cDNA末端的5倍放大 (5倍放大cDNA末端) 技术.
- 建设高收益,高质量的退化图书馆.
- 使用短读下一代测序 (NGS) 进行片段分析.
主要成果:
- 降解体概况可以验证预测的微RNA介导裂变事件.
- 通过这种方法可以识别新的microRNA目标.
- 获得了对RNA处理和RNA结合蛋白的作用的洞察.
结论:
- 描述的降解组测序方法对于分析RNA降解是有效的.
- 这种方法对研究非模型生物中的RNA动态非常有益.
- 该协议为退化图书馆的构建和分析提供了一个强大的框架.
相关概念视频
Proteins: From Genes to Degradation
Within a biological system, the DNA encodes the RNA, and the nucleotide sequence in the RNA further defines the amino acid sequence in the protein. This is referred to as “The Central Dogma of Molecular Biology” - a term coined by Francis Crick. Central dogma is a firm principle in biology that defines the flow of genetic information within any life form. The two fundamental steps in central dogma are - transcription and translation.
Transcription is the synthesis of RNA molecules by RNA...
Transcription is the synthesis of RNA molecules by RNA...
Sanger Sequencing
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
Next-generation Sequencing
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
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...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Maxam-Gilbert Sequencing
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
Challenges of the Maxam-Gilbert Method
The...
Proteins: From Genes to Degradation
Within a biological system, the DNA encodes the RNA, and the nucleotide sequence in the RNA further defines the amino acid sequence in the protein. This is referred to as “The Central Dogma of Molecular Biology” - a term coined by Francis Crick. Central dogma is a firm principle in biology that defines the flow of genetic information within any life form. The two fundamental steps in central dogma are - transcription and translation.
Transcription is the synthesis of RNA molecules by RNA...
Transcription is the synthesis of RNA molecules by RNA...


