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相关概念视频

RNA-seq03:21

RNA-seq

9.9K
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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RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

6.3K
Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...
6.3K
Ribosome Profiling02:24

Ribosome 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...
3.5K

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相关实验视频

Updated: Jun 24, 2025

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA
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线性放大单链RNA衍生的转录组测序 (LAST-seq)

Jun Lyu1, Chongyi Chen1

  • 1Laboratory of Biochemistry and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.

Bio-protocol
|June 14, 2024
PubMed
概括
此摘要是机器生成的。

我们开发了LAST-seq,一种新的单细胞RNA测序方法. 它直接放大RNA,提高灵敏度和减少噪声,以便更好地分析基因表达.

关键词:
基因表达噪声 基因表达噪声在体外转录 In vitro转录线性放大线性放大线性放大线单细胞RNA测序的一个细胞.单链RNA模板是一个单链RNA模板转录性爆发是指转录性爆发.

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相关实验视频

Last Updated: Jun 24, 2025

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14:49

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Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing
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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:

  • 单细胞RNA测序 (scRNA-seq) 对于生物和生物医学研究至关重要.
  • 目前的scRNA-seq方法使用逆转录 (RT) 和第二链合成 (SSS),通常限制检测灵敏度.
  • 在RT/SSS中缺乏效率会损害单细胞RNA的准确量化.

研究的目的:

  • 开发一种具有增强灵敏度和降低技术噪音的新型scRNA-seq方法.
  • 克服传统RT/SSS依赖的scRNA-seq协议的局限性.
  • 为了能够在单细胞水平上更精确地分析基因表达.

主要方法:

  • 开发了线性放大单链RNA衍生转录组测序 (LAST-seq).
  • LAST-seq直接放大单链RNA,绕过了RT和SSS的需要.
  • 利用RNase H/DNA聚合酶策略将T7促进体附加到单链RNA中,随后进行T7促进体介导的体外转录 (IVT).

主要成果:

  • 在单细胞转录组分析中,LAST-seq证明了高灵敏度RNA检测.
  • 与现有技术相比,该方法实现了低水平的技术噪声.
  • 成功地应用了LAST-seq来量化人类细胞中转录突破动力学.

结论:

  • 在单细胞RNA测序技术中,LAST-seq提供了显著的进步.
  • 该方法为转录组分析提供了更敏感,更低噪音的方法.
  • LAST-seq促进了对基因表达调节的更深入的了解,包括染色体组织的作用.