相关实验视频

Updated: May 16, 2025

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons

Published on: August 29, 2014

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 Graham^1,2, Fadheela Patel³, Francesca Little⁴

¹Imaging Sciences, Neuroscience Institute, University of Cape Town, Cape Town, South Africa.

Frontiers in bioinformatics

|April 1, 2025

概括

新的短读测序方法使得精确的物种级微生物组分析成为可能. 然而,采样方法对结果有显著的影响,因此在婴儿肠道微生物组研究中需要仔细考虑.

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核糖体RNA (rRNA) 基因的短读测序通常使用有限的可变区域,阻碍了物种级别的解析.
长读测序提供了完整的16S rRNA基因覆盖,但短读方法仍然很普遍.
新的套件和管道旨在通过短读测序来提高物种级别的识别和可重复性.

研究的目的:

评估精确的物种级别分辨率和可重复性的可行性,使用一种新的短读测序套件和生物信息学管道.
评估不同样本采集方法对婴儿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基因的所有九个变量区域进行测序.
包括模拟社区和复制样本用于质量控制和可重复性评估.
来自婴儿的配对便和直肠拭子样本的比较,分析微生物概况,使用统计测试来测量多样性.

主要成果:

该协议使用多个16S rRNA基因变量区域实现了精确的物种级别识别和高可重复性.
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.
模拟社区分析证实了物种级别分配的准确性.

结论:

开发的协议对于婴儿肠道微生物群落的物种级分析是有效的.
采样方法带来了变化,在下游数据分析中必须仔细考虑.
在解释婴儿微生物组数据时,未来的研究应该考虑样本类型的差异.

关键词:

16S rRNA 测序的测序方法微生物组是一个微生物组.多个变量区域是多个变量区域.简短阅读简短阅读在物种级别的物种级别.

更多相关视频

Efficient Nucleic Acid Extraction and 16S rRNA Gene Sequencing for Bacterial Community Characterization

Efficient Nucleic Acid Extraction and 16S rRNA Gene Sequencing for Bacterial Community Characterization

Published on: April 14, 2016

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing

Published on: October 15, 2019

相关概念视频

RNA-seq

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...

Gene Evolution - Fast or Slow?

Gene Evolution - Fast or Slow?

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...

Next-generation Sequencing

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....