.png){kind=icon}
返回
High-throughput microfluidic single-cell RT-qPCR
.png){kind=icon}
OA
AI
DOI：10.1073/pnas.1019446108.png)
.png){kind=icon}
原文求助.png){kind=icon}
分享.png){kind=icon}
收藏
摘要
En 中文
A long-sought milestone in microfluidics research has been the development of integrated technology for scalable analysis of transcription in single cells. Here we present a fully integrated microfluidic device capable of performing high-precision RT-qPCR measurements of gene expression from hundreds of single cells per run. Our device executes all steps of single-cell processing, including cell capture, cell lysis, reverse transcription, and quantitative PCR. In addition to higher throughput and reduced cost, we show that nanoliter volume processing reduced measurement noise, increased sensitivity, and provided single nucleotide specificity. We apply this technology to 3,300 single-cell measurements of (i) miRNA expression in K562 cells, (ii) coregulation of a miRNA and one of its target transcripts during differentiation in embryonic stem cells, and (iii) single nucleotide variant detection in primary lobular breast cancer cells. The core functionality established here provides the foundation from which a variety of on-chip single-cell transcription analyses will be developed.
Keyword:
real-time PCR
lab on a chip
transcriptional noise
microRNA
single-cell diagnostics
期刊
P
IF:
9.1
论文数:
10.8W
被引数:
73.5W
机构
引用论文
MicroRNA expression profiling of single whole embryonic stem cells单个完整胚胎干细胞MicroRNA表达谱分析.png)
NUCLEIC ACIDS RESEARCH
OAAI
NUCLEIC ACIDS RESEARCHIF13.1
OAAI分享收藏Resolution of Cell Fate Decisions Revealed by Single-Cell Gene Expression Analysis from Zygote to BlastocystDEVELOPMENTAL CELLOAAI
DEVELOPMENTAL CELLIF8.7
OAAI分享收藏Characterization of human embryonic stem cell lines by the International Stem Cell InitiativeNATURE BIOTECHNOLOGY
NATURE BIOTECHNOLOGYIF41.7
PREAI
分享收藏Excitatory and Inhibitory Effects of Serotonin on Sensorimotor Reactivity Measured with Acoustic StartleScience
ScienceIF0
PREAI
分享收藏Gene expression profiling in single cells from the pancreatic islets of Langerhans reveals lognormal distribution of mRNA levelsGENOME RESEARCHOAAI
GENOME RESEARCHIF5.5
OAAI分享收藏
查看更多内容