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Updated: Feb 17, 2026

Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

Published on: January 7, 2020

Multiplexing for Oxidative Bisulfite Sequencing (oxBS-seq).

Kristina Kirschner^1,2,3, Felix Krueger⁴, Anthony R Green^1,2,3,5

¹Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK.

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

|December 11, 2017

Summary

This summary is machine-generated.

This study introduces an improved oxidative Bisulfite Sequencing (oxBS-seq) method. It efficiently identifies 5-hydroxymethylcytosine (5hmC) genome-wide with reduced sample input and cost.

Keywords:

5-Hydroxymethylcytosine DNA Methylation Multiplexing Oxidative bisulfite sequencing Pooling

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Area of Science:

Epigenetics and Genomics
Molecular Biology
Biotechnology

Background:

DNA modifications, particularly DNA methylation, are vital for cellular regulatory processes.
5-hydroxymethylcytosine (5hmC) is a recently discovered DNA modification, an intermediate in 5-methylcytosine (5mC) oxidation.
Understanding the function of 5hmC is an active area of research, with several sequencing-based methods developed to quantify its levels.

Purpose of the Study:

To develop a more efficient and cost-effective method for genome-wide 5hmC detection.
To reduce the amount of starting DNA required for 5hmC analysis.
To facilitate broader research into the functional roles of 5hmC.

Main Methods:

Integration of adapter-based multiplexing into the oxidative Bisulfite Sequencing (oxBS-seq) workflow.
Incorporation of high-efficiency library preparation techniques.
Genome-wide assessment of 5-hydroxymethylcytosine (5hmC) levels.

Main Results:

Successfully reduced the required starting amount of DNA per sample.
Significantly lowered the cost per sample for 5hmC analysis.
Enabled efficient genome-wide identification of 5hmC levels.

Conclusions:

The adapted oxBS-seq workflow provides a more accessible and economical approach for genome-wide 5hmC profiling.
This method supports further investigation into the biological significance of 5hmC.
The reduced cost and input requirements democratize 5hmC research.