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Related Experiment Video

Updated: May 6, 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

Direct cell lysis for single-cell gene expression profiling.

David Svec¹, Daniel Andersson, Milos Pekny

¹Institute of Biotechnology AS CR , Prague , Czech Republic ; TATAA Biocenter , Gothenburg , Sweden.

Frontiers in Oncology

|November 14, 2013

Summary

This summary is machine-generated.

Direct cell lysis using bovine serum albumin (BSA) offers superior transcript yield and RNA stability for analyzing small cell samples. This method outperforms standard extraction techniques for single-cell and few-cell analyses.

Keywords:

DNA spike RNA purification RNA spike cell lysis direct lysis real-time PCR single-cell biology single-cell gene expression

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Area of Science:

Molecular Biology
Genomics
Biochemistry

Background:

Analyzing nucleic acids from single or few cells presents challenges due to low transcript numbers.
Existing extraction protocols often sacrifice yield for purity, limiting their use for minute samples.

Purpose of the Study:

To evaluate direct cell lysis protocols for nucleic acid extraction from small cell samples.
To determine the optimal lysis agent for maximizing transcript yield and downstream assay compatibility.

Main Methods:

Seventeen direct cell lysis protocols were assessed for transcript yield and suitability for reverse transcription quantitative real-time PCR (RT-qPCR).
Assays included four endogenous genes and RNA/DNA spikes across samples ranging from 1 to 512 mammalian cells.
Bovine serum albumin (BSA) was investigated as a key lysis agent.

Main Results:

Bovine serum albumin (BSA) demonstrated superior performance as a lysis agent, ensuring efficient cell lysis and high RNA stability.
BSA-based lysis enhanced reverse transcription efficiency compared to other agents.
Direct cell lysis with BSA proved more effective than standard column-based extraction for samples of 1-512 cells.

Conclusions:

Direct cell lysis protocols utilizing BSA are effective for small cell sample analysis.
BSA-based lysis is compatible with various cell collection methods and downstream analytical workflows.
This approach significantly improves transcript yield and assay performance for single-cell and few-cell studies.