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Next-generation Sequencing

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.

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Updated: May 24, 2026

Flow-sorting and Exome Sequencing of the Reed-Sternberg Cells of Classical Hodgkin Lymphoma

Flow-sorting and Exome Sequencing of the Reed-Sternberg Cells of Classical Hodgkin Lymphoma

Published on: June 10, 2017

Rapid flow-sorting to simultaneously resolve multiplex massively parallel sequencing products.

Julia Sandberg¹, Beata Werne, Mark Dessing

¹Science for Life Laboratory, Department of Gene Technology, Royal Institute of Technology (KTH), Box 1031, SE-17121, Solna, Sweden.

Scientific Reports

|February 23, 2012

Summary

This summary is machine-generated.

This study introduces a new method for normalizing multiplex next-generation sequencing libraries after emulsion PCR. Fluorescent tagging and flow cytometry sorting improve sequence quality and read length by removing unwanted beads.

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Published on: June 10, 2017

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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

Area of Science:

Molecular Biology
Genomics
Biotechnology

Background:

Next-generation sequencing (NGS) library preparation often involves bead-based amplification.
Multiplexing libraries enhances sequencing output but requires effective normalization.
Current enrichment protocols can lead to suboptimal sequence quality and read length.

Purpose of the Study:

To develop a novel method for normalizing multiplex NGS libraries post-emulsion PCR.
To improve sequence quality and mean read length in multiplex sequencing runs.
To provide a simple and effective alternative to standard enrichment protocols.

Main Methods:

Amplification of barcoded library fragments on bead surfaces.
Fluorescent tagging of amplified library fragments with complementary sequences.
High-speed flow cytometric sorting of fluorescently labeled emulsion PCR beads.

Main Results:

Achieved even sequence distribution of multiplex libraries after flow sorting.
Significantly increased sequence quality and mean read length.
Effectively removed empty and mixed emulsion PCR beads, enhancing data output.

Conclusions:

The novel fluorescent tagging and flow sorting method offers efficient normalization for multiplex NGS libraries.
This approach enhances sequencing data quality and yield compared to traditional methods.
The protocol is simple and applicable to various NGS platforms requiring bead-based amplification.