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Updated: Apr 11, 2026

Transcriptome Analysis of Single Cells

Transcriptome Analysis of Single Cells

Published on: April 25, 2011

Single prokaryotic cell isolation and total transcript amplification protocol for transcriptomic analysis.

Yun Kang¹, Ian McMillan², Michael H Norris³

¹Department of Microbiology, University of Hawaii at Manoa, Honolulu, Hawaii, USA.

Nature Protocols

|June 5, 2015

Summary

This summary is machine-generated.

Single prokaryotic cell transcriptome analysis is now possible. This new method amplifies total RNA for detailed gene expression insights using laser capture and enzymatic amplification.

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Last Updated: Apr 11, 2026

Transcriptome Analysis of Single Cells

Transcriptome Analysis of Single Cells

Published on: April 25, 2011

Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

Published on: January 16, 2017

An Oligonucleotide-based Tandem RNA Isolation Procedure to Recover Eukaryotic mRNA-Protein Complexes

An Oligonucleotide-based Tandem RNA Isolation Procedure to Recover Eukaryotic mRNA-Protein Complexes

Published on: August 18, 2018

Area of Science:

Microbiology
Molecular Biology
Genomics

Background:

Single-cell transcriptome analysis offers detailed spatiotemporal gene expression insights.
Previous methods were limited to eukaryotic cells, excluding prokaryotes.
Expanding single-cell transcriptomics to prokaryotes is crucial for comprehensive biological understanding.

Purpose of the Study:

To develop and describe a robust procedure for analyzing the transcriptome of single prokaryotic cells.
To enable in-depth gene expression profiling at the single-cell level in prokaryotes.
To facilitate the application of advanced molecular techniques to prokaryotic single-cell research.

Main Methods:

Single-cell isolation using laser-capture microdissection.
RNA reverse transcription, DNA digestion, and cDNA amplification using specific enzymes (Moloney murine leukemia virus, McrBC, DpnI, T4 polynucleotide kinase, CircLigase, Φ29 polymerase).
A 5-day protocol yielding double-stranded cDNA (ds-cDNA) suitable for microarray analysis.

Main Results:

Successful amplification of total RNA from individual prokaryotic cells.
Generation of sufficient ds-cDNA for downstream applications like microarray analysis.
Demonstration of a feasible workflow for single prokaryotic cell transcriptome analysis.

Conclusions:

The described procedure effectively enables transcriptome analysis of single prokaryotic cells.
This advancement opens new avenues for studying prokaryotic gene expression heterogeneity.
The method provides a valuable tool for detailed molecular investigations in microbiology.