使用牛津纳米孔小ION进行完整细菌基因组测序和组装的逐步指南
Anil Kumar1, Max M Häggblom1, Lee J Kerkhof2
1Department of Biochemistry and Microbiology, Rutgers University, New Brunswick, NJ, USA.
Methods in molecular biology (Clifton, N.J.)
|November 15, 2024
概括
牛津纳米孔 (ONT) MinION测序为完整的微生物基因组组装提供了长读数. 这种方法使用R9.4流细胞和多拉多软件,提供高质量,具有成本效益的细菌基因组生成.
科学领域:
- 基因组学就是基因组学.
- 微生物学 微生物学
- 生物信息学是一种生物信息学.
背景情况:
- 传统的测序方法在生成完整的微生物基因组方面存在局限性.
- 牛津纳米孔 (ONT) 技术提供长读测序能力.
- 在ONT R9.4流量电池和多拉多软件的进步提高了读取精度.
研究的目的:
- 用ONT MinION描述一种详细的方法来完成微生物基因组组装.
- 概述了抛光和评估组装的基因组质量的步骤.
- 突出ONT MinION在经济高效的微生物基因组生成中的实用性.
主要方法:
- 使用牛津纳米孔 (ONT) MinION用于长读DNA/RNA测序.
- 采用R9.4流量单元来提高测序精度.
- 应用多拉多软件用于基础调用和读取生成.
- 使用各种生物信息学工具进行基因组组装,抛光和质量评估.
主要成果:
- 产生高质量的长读物,适用于完整的细菌基因组组装.
- 成功地组装和抛光微生物基因组.
- 证明ONT MinION在生产高质量的基因组数据方面的有效性.
结论:
- 使用R9.4流细胞和多拉多软件的ONT MinION平台,可实现高质量,完整的微生物基因组组装.
- 这种具有成本效益的方法可用于寻求生成微生物基因组的实验室.
- 描述的协议有助于对各种微生物进行强大的基因组分析.
相关概念视频
Genome Annotation and Assembly
18.8K
The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.
18.8K
RNA-seq
9.8K
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...
9.8K
Sanger Sequencing
753.0K
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...
753.0K
Next-generation Sequencing
87.6K
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....
87.6K


