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RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...

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Updated: Jul 12, 2026

Microfluidics-based High-throughput Circulating Tumor Cell Sorting and Single-cell Sequencing Technology

Microfluidics-based High-throughput Circulating Tumor Cell Sorting and Single-cell Sequencing Technology

Published on: November 14, 2025

Micro RNA Sequencing in Circulating Tumor Cells.

Sarah M Hücker¹, Stefan Kirsch²

¹Fraunhofer Institute for Toxicology and Experimental Medicine, Division Personalized Tumor Therapy, Am Biopark 9, 93053, Regensburg, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|July 9, 2026

Summary

This study presents a protocol for preparing microRNA (miRNA) sequencing libraries from circulating tumor cells (CTCs). This method enables accurate miRNA quantification at the single-cell level, crucial for cancer diagnostics and personalized medicine.

Keywords:

Circulating tumor cell NGS Single-cell sequencing Small RNA microRNA

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Last Updated: Jul 12, 2026

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Published on: March 14, 2019

Area of Science:

Oncology
Molecular Biology
Genomics

Background:

MicroRNAs (miRNAs) are key regulators in cancer biology and potential biomarkers for prognosis and diagnosis.
Circulating tumor cells (CTCs) harbor miRNAs that can indicate early metastasis, drug resistance, and disease recurrence.
Accurate miRNA quantification in single CTCs is essential for exploiting their biomarker potential.

Purpose of the Study:

To develop and describe a protocol for miRNA sequencing library preparation specifically for CTCs.
To enable accurate miRNA expression profiling at the single CTC level.

Main Methods:

The protocol involves cell lysis, adapter ligation to miRNAs, reduction of non-target RNA and adapter dimers via exonuclease digestion, reverse transcription, amplification, and purification.
It is compatible with various single-cell isolation techniques like micromanipulation and FACS sorting.
Quality control steps are included to ensure high-quality samples for sequencing.

Main Results:

A robust protocol for miRNA library preparation from CTCs was established.
The method allows for sensitive and accurate miRNA quantification from limited cell numbers.
Quality control metrics were defined for sample selection.

Conclusions:

The described protocol facilitates the detailed analysis of miRNA expression in single CTCs.
This advancement supports the use of miRNAs as biomarkers for early cancer detection, treatment response prediction, and monitoring disease progression.
Accurate single-cell miRNA profiling in CTCs holds significant potential for personalized cancer therapy.