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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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Next-generation Sequencing03:00

Next-generation Sequencing

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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....
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Sanger Sequencing01:57

Sanger Sequencing

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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...
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Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

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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
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RACE - Rapid Amplification of cDNA Ends02:35

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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...
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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
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Updated: Jan 17, 2026

Amplicon Sequencing using the Long-Read Sequencing Technologies
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Amplicon Sequencing using the Long-Read Sequencing Technologies

Published on: August 29, 2025

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使用长读测序技术进行幅测序.

Morwasehla Modjadji1, Brendon Coenrad Mann1, Johannes Loubser1

  • 1Department of Science and Innovation - National Research Foundation Centre of Excellence for Biomedical Tuberculosis Research, South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University.

Journal of visualized experiments : JoVE
|September 15, 2025
PubMed
概括
此摘要是机器生成的。

便携式测序为检测耐药结核病 (DR-TB) 提供了一种具有成本效益的方法. 这种方法可以准确识别高频耐药性突变,但可能会错过低频变异,影响资源有限的环境中的结核病诊断.

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相关实验视频

Last Updated: Jan 17, 2026

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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.
  • 传染病诊断 传染病诊断 传染病诊断

背景情况:

  • 世界卫生组织 (WHO) 强调需要可访问的结核病 (TB) 诊断.
  • 下一代测序 (NGS) 提高了耐药结核病 (DR-TB) 的检测,有针对性的NGS (tNGS) 专注于耐药性突变.
  • 高成本的测序平台限制了低收入和中等收入国家的DR-TB诊断.

研究的目的:

  • 用tNGS试验评估便携式测序对于DR-TB检测的实用性.
  • 为了比较便携式 (长读) 测序与高精度 (短读) 测序的性能,用于识别结核病中与耐药性相关的变体.

主要方法:

  • 从耐利芬素结核病 (RR-TB) 隔离物的DNA使用tNGS试验进行了放大.
  • 放大产品在便携式测序平台和高精度短读平台上进行了测序.
  • 生物信息学管道用于长读和短读数据被用于分析.

主要成果:

  • 便携式测序方法成功识别了与高频电阻相关的变体.
  • 便携式测序的结果与常见变体的参考短读测序平台的相关性很好.
  • 在使用便携式测序方法检测低频电阻相关变异时观察到局限性.

结论:

  • 便携式测序是DR-TB诊断的可行,具有成本效益的替代方案,特别是在资源有限的环境中.
  • 虽然对高频变体有效,但需要进一步优化,以提高对低频变体在结核病耐药性检测中的灵敏度.