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Updated: Jun 10, 2026

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Published on: February 25, 2017

Amplification-free digital gene expression profiling from minute cell quantities.

Fatih Ozsolak¹, David T Ting, Ben S Wittner

¹Helicos BioSciences Corporation, Cambridge, Massachusetts, USA. fatihozsolak@gmail.com

|July 20, 2010

Summary

This summary is machine-generated.

This study introduces low-quantity digital gene expression (LQ-DGE), an amplification-free method for gene profiling from small cell samples. LQ-DGE enables accurate gene expression analysis, even from limited or archival cell materials.

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Area of Science:

Molecular Biology
Genomics
Biotechnology

Background:

Accurate gene expression profiling is essential for scientific research and clinical diagnostics.
Existing methods often require substantial cell quantities, limiting their application.
Minute cell samples pose challenges for reliable gene expression analysis.

Purpose of the Study:

To develop an amplification-free method for gene expression profiling from minute cell quantities.
To demonstrate the quantitative accuracy and broad applicability of the new method.
To enable gene expression analysis in scenarios with limited cellular material.

Main Methods:

Low-Quantity Digital Gene Expression (LQ-DGE) captures poly(A)(+) RNAs from cell lysates.
RNA is immobilized on poly(dT)-coated sequencing surfaces.
On-surface reverse transcription and sequencing are performed without amplification.

Main Results:

LQ-DGE successfully generated reliable expression profiles from minute cell samples.
The method demonstrated quantitative accuracy in profiling both malignant and nonmalignant cells.
Application to mouse and human cells confirmed the technique's versatility.

Conclusions:

LQ-DGE is a powerful, amplification-free approach for gene expression profiling from minimal cell inputs.
This technique has significant potential for scientific discovery and clinical applications, including archival specimens.
LQ-DGE overcomes limitations of existing methods when dealing with scarce cellular material.