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Related Experiment Video

Updated: Apr 11, 2026

Sequencing of mRNA from Whole Blood using Nanopore Sequencing

Sequencing of mRNA from Whole Blood using Nanopore Sequencing

Published on: June 3, 2019

Phasing of single DNA molecules by massively parallel barcoding.

Erik Borgström¹, David Redin¹, Sverker Lundin¹

¹Science for Life Laboratory, Division of Gene Technology, School of Biotechnology, Royal Institute of Technology (KTH), SE-171 65 Solna, Sweden.

Nature Communications

|June 10, 2015

Summary

This summary is machine-generated.

This study introduces a novel method for barcoding and phasing single DNA molecules, preserving crucial genetic information lost in short-read sequencing. This technique allows for the analysis of long DNA molecules on standard platforms, enhancing variant phasing accuracy.

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Area of Science:

Genomics
Molecular Biology
Bioinformatics

Background:

High-throughput sequencing often yields short reads, leading to loss of genetic variant phasing information.
Determining which alleles are linked on individual DNA molecules is critical for various genetic analyses.
Existing methods may not efficiently recover phase information from short-read data.

Purpose of the Study:

To develop and validate a method for massively parallel barcoding and phasing of single DNA molecules.
To enable the analysis of long DNA molecules and their associated genetic variants using short-read sequencing platforms.
To recover lost phasing information crucial for understanding genetic variation.

Main Methods:

Generation of a primer library with millions of uniquely barcoded beads.
Compartmentalization of beads with single DNA molecules for amplification and tagging.
Application of the assay to bacterial 16S sequencing for validation.

Main Results:

Demonstrated a method for massively parallel barcoding and phasing of single DNA molecules.
Successfully coupled biological information from multiple loci on single DNA molecules.
Achieved up to 94% of hypothesized phasing events originating from single molecules in 16S sequencing.

Conclusions:

The developed method effectively enables the study of long single molecules within complex samples using short-read sequencing.
Phase information, crucial for genetic variant analysis, can be retained despite using short-read platforms.
This approach significantly improves the utility of widely available sequencing technologies for comprehensive genetic analysis.