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Updated: Apr 7, 2026
07:27
Transcriptome Analysis of Single Cells
Published on: April 25, 2011
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Whole-Transcriptome Amplification of Single Cells for Next-Generation Sequencing
Christian Korfhage1, Evelyn Fricke1, Andreas Meier1
1QIAGEN GmbH, Hilden, Germany.
Current Protocols in Molecular Biology
|July 2, 2015
Summary
Single-cell transcriptome analysis reveals cell variability in health and disease. Whole-transcriptome amplification (WTA) enables next-generation sequencing (NGS) of single cells, providing crucial biological insights.
Area of Science:
- Molecular Biology
- Genomics
- Cell Biology
Background:
- Single-cell transcriptome analysis offers insights into cellular heterogeneity and its role in biological functions.
- Cellular variability can serve as an indicator of disease states, such as cancer.
- Next-generation sequencing (NGS) platforms are advancing the study of cell variability.
Purpose of the Study:
- To describe a comprehensive workflow for single-cell transcriptome analysis using NGS.
- To detail the process from cell preparation to NGS library construction.
- To provide quality control recommendations for whole-transcriptome amplification (WTA).
Main Methods:
- Cell quality testing
- Cell lysis and gDNA removal
- Whole-transcriptome amplification (WTA)
- NGS library preparation
Main Results:
- A complete workflow for single-cell transcriptome analysis is presented.
- Quality control measures for WTA are recommended.
- Optional protocols for RNA purification and WTA purification are included.
Conclusions:
- Accurate whole-transcriptome amplification is essential for analyzing single-cell transcriptomes via NGS.
- The described workflow facilitates the elucidation of cell variability in both healthy and diseased states.
- This methodology supports deeper understanding of cellular heterogeneity and its implications.
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