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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...

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Related Experiment Video

Updated: Jun 23, 2025

G2-seq: A High Throughput Sequencing-based Technique for Identifying Late Replicating Regions of the Genome

G2-seq: A High Throughput Sequencing-based Technique for Identifying Late Replicating Regions of the Genome

Published on: March 22, 2018

Quantifying DNA replication speeds in single cells by scEdU-seq.

Jeroen van den Berg¹, Vincent van Batenburg², Christoph Geisenberger^2,3

¹Oncode Institute, Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences) and University Medical Center Utrecht, Utrecht, The Netherlands. j.berg@hubrecht.eu.

|June 17, 2024

Summary

This summary is machine-generated.

DNA replication speed accelerates during the S phase of the cell cycle. Transcription-induced DNA damage limits early replication speed, while later replication in less-transcribed regions speeds up.

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Last Updated: Jun 23, 2025

G2-seq: A High Throughput Sequencing-based Technique for Identifying Late Replicating Regions of the Genome

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Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

DNA replication is essential for cell division, involving thousands of replication forks.
Replication rate may be influenced by epigenetic factors and cell type.
Accurate measurement of DNA replication speeds is crucial for understanding genome duplication.

Purpose of the Study:

To develop and apply a method for measuring DNA replication speeds in single cells.
To investigate the dynamics of DNA replication rate throughout the S phase.
To identify factors influencing the acceleration of DNA replication.

Main Methods:

Development of single-cell 5-ethynyl-2'-deoxyuridine sequencing (seqEdU-seq).
Detection of nascent replicated DNA using seqEdU-seq.
Application of genetic and pharmacological perturbations to study replication dynamics.

Main Results:

DNA replication speed is not constant and increases during S phase.
Transcription-induced DNA damage limits replication speed in early S phase.
Replication accelerates in late S phase as less-transcribed regions are replicated.

Conclusions:

Replication fork speed is regulated during the cell cycle.
Transcription-associated DNA damage is a key factor limiting replication speed.
Understanding replication dynamics is vital for genome stability and cell proliferation.