The Microbiome Blog | Cmbio

What is High Throughput Sequencing?

Written by Manoj Dadlani | Mar 6, 2025 11:30:12 AM

 

 

The use of high throughput sequencing in metagenomics is becoming increasingly important as researchers strive to study the interactions between different microorganisms and their environment. High-throughput sequencing has now revolutionised almost all areas of the scientific field. But what is high throughput sequencing, how does it work, and what are its applications?

In this guide, we will discuss the types of high throughput sequencing technologies that are available, their applications in metagenomics, and some of the basics of using these methods. Let's dive in.

 

So - What is High Thoughput Sequencing?

High throughput sequencing (also known as next generation sequencing) is a way to study the interactions between different microorganisms and their environment. It involves the use of technologies that can sequence vast amounts of DNA quickly, and has revolutionized the sequencing world.

Organizations like the Human Genome Project have used this technology to yield 90% of the human genome sequence in draft form in order to learn more about the entire human genome, whole genome sequencing, diseases and how they are inherited.

This technology is becoming more and more important as researchers strive to learn more about gene expression and how microorganisms interact with each other.

How does high throughput genome sequencing work?

Sequencing DNA is like taking a picture of it, and next generation sequencing uses machines that can take numerous pictures of DNA molecules very quickly. This not only allows researchers to study a lot of DNA at the same time but it also allows them to sequence data on a much larger scale.

One example of how high throughput sequencing works in metagenomics is in the study of microbial communities. Next generation genome sequences take numerous 'snapshots' of the DNA present in a microbial community, providing deep insights into the interactions between different microorganisms.

By using these techniques, researchers can better understand the microbiome, biodiversity and interactions between different microorganisms at scale, which can help us to identify new species and predict how they will interact with each other and their environment.

High throughput sequencing technology

There are a few different high-throughput genome sequencing technologies that are available, each of which has its own advantages and disadvantages. Some different types of high-throughput DNA sequencing technology include:

  1. Sequencing by synthesis: This technology uses machines that can sequence genomic DNA very quickly. It is one of the most popular high throughput sequencing technologies because it is very efficient and can produce a lot of data, including specific sequencing technologies such as Illumina and PacBio. 
  2. Sequencing by unfolding: Other popular technologies like Oxford Nanopore Technology (ONT) read the bases of a DNA strand as it moves through the nanopore protein.

3 Types of High Throughput Sequencing

There are a few different types of high throughput sequencing technologies, each of which has its own advantages and disadvantages. These technologies include:

  • Bridge amplification: This type of sequencing is based in a flow cell, which produces clusters of DNA strands for further sequencing and analysis. The flow cell has two oligos that are complementary to the adapters on the fragment strand and can thus be used to produce more copies of the DNA sequence.
  • Single molecular real-time sequencing: This single cell sequencing method can sequence DNA very quickly using a parallelized single molecule DNA sequencing method. Single molecule sequencing enables direct observation of a single molecule or entire genome in real time.
  • Nanopore-based sequencing: This type of high throughput sequencing employs a protein nanopore that detects changes in electric current as nucleic acids travel through it. It is a scalable and unique technology that enables direct, real-time analysis of RNA sequencing fragments or long DNA.

4 Advantages of High Throughput Sequencing

High throughput sequencing technologies offer a number of advantages over other sequencing methods. These advantages include:

  • Faster data generation rates: High throughput sequencing technologies can generate data much more quickly than low throughput sequencing technologies. This makes them ideal for applications that require large amounts of data, such as metagenomics.
  • Better accuracy and dynamic range: High throughput sequencing technologies are more accurate than low throughput sequencing technologies. This means that they are better able to produce accurate results, even when sequencing data from complex DNA samples.
  • Detailed insights into cellular genomics and transcriptomics: Since high throughput sequencing enables direct observation of single nucleotide variants in real time, it can provide deep insights into cellular genomics and transcriptomics. This makes it an important tool for researchers studying molecular biology and the human genome.
  • Large data sets: High throughput sequencing technologies can produce large data sets that enable researchers to gain a more detailed understanding of cellular processes and gene expression in clinical samples. This makes them especially useful for applications in the study of viral genomes, in drug discovery and in disease research.
 

Key High Throughput Sequencing Applications

 

Biomedical research

One of the key high throughput sequencing applications is biomedical research. By providing researchers with detailed insights into cellular genomics and transcriptomics, high throughput sequencing helps them better understand complex biological processes and disease pathways.

Genomic and transcriptomic signatures of various diseases and developmental stages can be used to:

  1. Characterize biological markers (biomarkers) including genes and proteins often derived from human tissue or blood. Biomarkers can be classified into three categories: diagnostic (presence or absence of disease), predictive (how a patient responds to treatment) and prognostic (how long a patient survives post-intervention), aiding in the development of new therapies and treatments.
  2. Understand disease development and progression and/or predict treatment response or patient survival. These insights can also help improve vaccines, drugs, and other interventions that target specific pathways in the cell or genome.

By analyzing genomic and transcriptomic signatures, researchers are able to better understand the molecular mechanisms underlying disease development, progression, and response to treatment. This enables them to more effectively develop new therapies for a wide range of diseases, from cancer to neurological disorders.

Clinical application

High throughput sequencing is also widely used in clinical research, as it enables researchers to identify biomarkers and better understand disease pathways. This information can then be used to develop new diagnostic tools and improve patient outcomes.

For example, high throughput methods can be used in in-vitro diagnostics, which target the region around the CFTR gene at the genomic level, for screening and diagnosis purposes. By identifying mutations in this gene, researchers can develop targeted treatments for diseases like cystic fibrosis.

Similarly, high throughput sequencing has been used to identify biomarkers and improve our understanding of various other diseases, including cancer, heart disease, diabetes, and Alzheimer's. By combining high throughput sequencing with other clinical technologies such as genetic testing and mass spectrometry, researchers are able to develop new tools for disease detection and monitoring, as well as personalized treatment approaches.

Forensic science

Another important application for high throughput sequencing is forensic science. In this field, high-throughput methods can be used to generate large amounts of data that can be used to identify individuals, estimate postmortem intervals and further investigate criminal cases.

For example, high throughput sequencing has been used to identify mutations in the genomes and transcriptomes of hair follicles. By analyzing these genetic markers, forensic scientists can obtain a detailed "genetic profile" for each individual, which they can then use to link individuals to crime scenes or exonerate suspects.

Additionally, they can use this information to identify individuals based on samples that may be too degraded for other forms of analysis, such as DNA samples.

Final Thoughts

The advantages of high throughput sequencing methods are many, and the applications are vast. High throughput sequencing is used in a wide range of research fields, from biomedical research to forensic science. Additionally, it has the potential to revolutionize diagnostics and personalized medicine.

Cmbio is a leader in metagenomics services and our methods provide researchers with unparalleled insights into the genomic and transcriptomic signatures of various diseases.

By leveraging high throughput sequencing methods, we help researchers identify biomarkers and better understand disease pathways. This information can then be used to develop new diagnostic tools and improve patient outcomes.

For more information on our sequencing and bioinformatics services

 

 
 

High Throughput Sequencing: FAQs

 

What's the difference between HTS and Sanger Sequencing?

High throughput sequencing methods are a type of next-generation sequencing (NGS) tool that generate large amounts of data at high speeds. In contrast, Sanger sequencing is a laboratory technique that relies on the use of specific enzymes to sequence DNA fragments in single or paired strands.

Can high-throughput sequencing execute RNA extraction?

Yes, high-throughput sequencing methods can be used to extract and sequence RNA. These methods offer many advantages over traditional RNA extraction techniques, including increased speed and accuracy.

Can high-throughput sequencing detect DNA polymerase?

Yes, high-throughput sequencing can be used to detect DNA polymerase activity. This is done by identifying the presence of specific DNA polymerase enzymes in a sample, which are typically associated with DNA replication and repair processes.