相关实验视频
Updated: Jun 25, 2025
10:24
Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
Published on: August 29, 2014
83.4K
使用多个可变区域的短读16SrRNA测序,以产生高质量的物种级结果
Amy S Graham1,2, Fadheela Patel3, Francesca Little4
1Imaging Sciences, Neuroscience Institute, University of Cape Town, Cape Town, South Africa.
bioRxiv : the preprint server for biology
|May 27, 2024
概括
新的短读测序方法可以准确地识别婴儿肠道微生物组中的物种. 然而,采样方法显著影响结果,需要在微生物组分析中仔细考虑.
科学领域:
- 微生物学 微生物学
- 基因组学就是基因组学.
- 生物信息学是一种生物信息学.
背景情况:
- 16S rRNA基因的短读测序通常针对有限的变量区域,限制物种级别的分辨率.
- 长读测序通过覆盖所有9个变量区域提供更高的分辨率,但成本更高.
- 精确的物种级微生物组表征对于理解健康和疾病至关重要.
研究的目的:
- 评估使用新型短读测序套件和生物信息管道实现精确的物种级别分辨率和可重复性的可行性.
- 评估不同样本采集方法 (便与直肠拭子) 对微生物组分析结果的影响.
主要方法:
- 在Illumina MiSeq平台上使用xGenTM 16S Amplicon Panel v2套件对16S rRNA基因的所有9个可变区域进行测序.
- 整合模拟社区和复制样本用于质量控制,并评估来自同一参与者的便和直肠拭子样本.
- 使用统计分析,包括威尔科克森等级总和测试和类内相关系数,以比较多样性指标和相对丰度.
主要成果:
- 该协议使得精确的物种级别识别和高度可复制的结果成为可能.
- 在同一个婴儿同时采集的便和直肠拭子样本之间的微生物概况中观察到显著差异.
- 这种新方法在物种层面描述婴儿肠道微生物群方面表现出有效性.
结论:
- 开发的协议是有效的物种层面分析婴儿肠道微生物组使用短读测序.
- 采样方法是一个关键因素,必须在微生物组数据的下游分析中加以解决.
- 进一步的研究应该考虑样本类型对微生物组研究结果的影响.
相关概念视频
02:05Gene Evolution - Fast or Slow?
7.1K
The genomes of eukaryotes are punctuated by long stretches of sequence which do not code for proteins or RNAs. Although some of these regions do contain crucial regulatory sequences, the vast majority of this DNA serves no known function. Typically, these regions of the genome are the ones in which the fastest change, in evolutionary terms, is observed, because there is typically little to no selection pressure acting on these regions to preserve their sequences.
In contrast, regions which code...
In contrast, regions which code...
7.1K
03:21RNA-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...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
9.9K
02:54Evolutionary Relationships through Genome Comparisons
5.7K
Genome comparison is one of the excellent ways to interpret the evolutionary relationships between organisms. The basic principle of genome comparison is that if two species share a common feature, it is likely encoded by the DNA sequence conserved between both species. The advent of genome sequencing technologies in the late 20th century enabled scientists to understand the concept of conservation of domains between species and helped them to deduce evolutionary relationships across diverse...
5.7K