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Next-generation Sequencing

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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.
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Related Experiment Video

Updated: Sep 10, 2025

Ultra-long Read Sequencing for Whole Genomic DNA Analysis

Ultra-long Read Sequencing for Whole Genomic DNA Analysis

Published on: March 15, 2019

High-throughput PacBio library preparation and sequencing techniques for genomic DNA and TNA.

Meghan L Bentz¹, Cristina Clines¹, Jasmine Padilla¹

¹Genomics Sequencing Laboratory, Centers for Disease Control and Prevention, Atlanta, GA, United States.

Frontiers in Genetics

|August 22, 2025

Summary

This summary is machine-generated.

This study presents two high-throughput methods for PacBio sequencing library preparation, enabling efficient whole genome assembly and analysis of clinical specimens. These workflows successfully generated thousands of genomic DNA and cDNA sequences.

Keywords:

COVID PacBio WGS whole-genome sequencing automation gDNA high throughput methods sequencing

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Last Updated: Sep 10, 2025

Ultra-long Read Sequencing for Whole Genomic DNA Analysis

Ultra-long Read Sequencing for Whole Genomic DNA Analysis

Published on: March 15, 2019

Ultralow Input Genome Sequencing Library Preparation from a Single Tardigrade Specimen

Ultralow Input Genome Sequencing Library Preparation from a Single Tardigrade Specimen

Published on: July 15, 2018

Simple and Rapid Method to Obtain High-quality Tumor DNA from Clinical-pathological Specimens Using Touch Imprint Cytology

Simple and Rapid Method to Obtain High-quality Tumor DNA from Clinical-pathological Specimens Using Touch Imprint Cytology

Published on: March 21, 2018

Area of Science:

Genomics and Bioinformatics
Molecular Biology and Genetics

Background:

PacBio Sequencing technology offers long read lengths crucial for closing gaps in whole genome assemblies.
There is a need for automated, high-throughput library preparation workflows for PacBio sequencing to enhance efficiency.

Purpose of the Study:

To describe two high-throughput methods for automated library preparation for PacBio sequencing.
To detail workflows for barcoded genomic DNA and cDNA library preparation, pooling, and sequencing on the Sequel II instrument.

Main Methods:

Development and implementation of two distinct high-throughput library preparation methods.
Barcoding of genomic DNA and cDNA samples for multiplexed sequencing.
Sequencing of prepared libraries using the PacBio Sequel II instrument.

Main Results:

Testing on 380 genomic DNA samples showed a 7.4% failure rate requiring repeat sequencing.
Testing on 384 Threose nucleic acid samples resulted in 0 failures, with 12.8% achieving <90% genome coverage.
The methods were successfully applied to generate approximately 2,000 genomic DNA and 30,000 cDNA genome sequences from clinical specimens in 2023.

Conclusions:

The described high-throughput methods are effective for PacBio sequencing library preparation.
These workflows facilitate large-scale genomic and transcriptomic sequencing from clinical samples.
The methods demonstrate scalability and utility for generating substantial genomic data.