测序:10X基因组学3'HT测试用于基因表达
1Molecular Biology & Genetics, Johns Hopkins School of Medicine, Baltimore, MD, USA. Linda.Orzolek@psomagen.com.
Methods in molecular biology (Clifton, N.J.)
|June 22, 2024
概括
10X Genomics 3'HT测定使单细胞RNA测序具有高通量,通过将细胞分离成滴并编码它们的cDNA. 这种先进的方法允许对成千上万个单个细胞进行详细的基因表达分析.
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) 对于理解细胞异质性至关重要.
- 现有的方法需要有效的细胞隔离和cDNA条形码,以进行高通量分析.
研究的目的:
- 描述10X基因组学3'高通量 (HT) 试验的方法.
- 详细介绍从细胞捕获到为scRNA-seq.q.准备库的过程.
主要方法:
- 采用以滴滴为基础的微流体系统,用于在油乳液中单细胞隔离.
- 采用具有独特细胞条形码的凝珠用于cDNA放大.
- 包含一个新的芯片设计,用于高容量的细胞捕获 (高达6万个细胞/车道) 和多样本处理 (高达16个样本/运行).
主要成果:
- 3'HT测定支持整个细胞和细胞核作为输入.
- 达到大约60%的细胞捕获率.
- 通过细胞条形编码在单个细胞中区分基因表达模式.
结论:
- 10X Genomics 3'HT测定为高通量单细胞基因表达概况提供了一个可扩展的解决方案.
- 描述的方法有助于样品质量控制,细胞捕获和下游测序的库准备.
- 这项技术提高了在前所未有的规模解决细胞异质性的能力.
相关概念视频
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...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
9.9K
DNA Microarrays
17.3K
Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
17.3K


