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Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells
Published on: January 7, 2020
Highly parallel genome-wide expression analysis of single mammalian cells
Jian-Bing Fan1, Jing Chen, Craig S April
1Research and Development, Illumina, Inc, San Diego, California, United States of America. jfan@illumina.com
Plos One
|February 21, 2012
Summary
We developed a high-throughput method for robust gene expression profiling from limited RNA. This technique enables sensitive whole-genome analysis in single cells or low-input samples.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Generating gene expression profiles typically requires significant RNA input.
- Analyzing gene expression in single cells or with limited RNA presents technical challenges.
Purpose of the Study:
- To develop a high-throughput amplification method for robust gene expression profiling.
- To enable accurate gene expression analysis from single-cell or low RNA inputs.
Main Methods:
- Utilized tagged priming and template-switching for cDNA synthesis.
- Incorporated universal PCR priming sites for global amplification.
- Integrated with a whole-genome gene expression microarray platform.
Main Results:
- Achieved high correlation values (R(2)~0.76-0.80) between individual cells.
- Demonstrated strong correlation (R(2)~0.69) between low RNA input replicates (50 pg).
- Detected approximately 63% of genes from single cells compared to 1 ng input.
Conclusions:
- Facilitates whole-genome gene expression profiling with extremely limited starting material.
- Applicable to studying progenitor cells in early development.
- Valuable for tumor stem cell biology research.
