Next Generation Sequencing: A Brief Guide to NGS Technology

The rise of Next Generation Sequencing has brought a huge transformative shift to genomics, making what was a lengthy and expensive process faster, more affordable, and more accessible. If you are interested in learning more about next-generation sequencing, this blog post will help you.

It has disrupted everything from clinical diagnostics and personalized medicine to agriculture and evolutionary biology. Let us discuss the core steps, including next generation sequencing, compasses, and the applications of NGS.

1. What is Next Generation Sequencing?

Next generation sequencing is a set of state-of-the-art DNA sequencing approaches that qualify for huge parallel sequencing of genetic material.

As well as Sanger sequencing, which sequences DNA one fragment at a time, another one, like NGS technologies, can significantly sequence millions to billions of fragments.

While comparing Sanger sequencing and next generation sequencing, the biggest difference is in scale, cost, and speed. Nowadays, platforms like Illumina’s next generation sequencing and Oxford Nanopore support researchers to sequence entire genomes within a few hours.

2. How Does NGS Work?

The next generation sequencing method begins with amino acid isolation, followed by library preparation. Also, the libraries are read on the Illumina sequencing system, which has been made to support a wide range of applications and thoughts.

Let us have a look at the workflow of NGS.

2.1 Nucleic acid Isolation: When extracting DNA from the cells or tissues, ensure optimal crop purity and quality.

2.2 Library Practice: Piece nucleic acids, connect adaptors and barcodes for multiplexing, and optionally function enrichment for targeted sequencing.

2.3 Sequencing: Utilize a sequencing platform such as Illumina to generate sequence data.2

2.4 Data Analysis: The process of sequencing reads, the quality control steps, and performs downstream analyses such as alignment, variant calling, and annotation.

3. Types of Next Generation Sequencing Techniques

Let us have a detailed look at the different types of generation sequencing techniques and their functionalities one by one here.

1️⃣ Sequencing By Synthesis – Illumina

This widely worn method employs reversible terminators to detect the incorporation of new nucleotides at the time of DNA synthesis.

Applications:

❶ Whole-genome sequencing
❷ Exome Sequencing
❸ RNA sequencing         
❹ Metagenomics
❺ Epigenomics, etc.

2️⃣ Sequencing by Migration

This next generation DNA sequencing through the Migration process utilizes labeled oligonucleotides that bind to the DNA template and ligate them to adjacent bases, which are the real. Meanwhile, this case involves the use of two base encoders to achieve more accuracy.

Applications:

❶ Genome-wide studies
❷ SNP detection
❸ Exome sequencing
❹ Comparative genomics

3️⃣ Pyrosequencing

It may detect the incorporation of a nucleotide by calculating the release of pyrophosphate when a nucleotide is supplemental to the growing DNA strand. The pyrophosphate is then converted to a light signal, which is consistent in real-time.

Applications

❶ Variant detection
❷ Targeted sequencing
❸ Genomic research
❹ SNP analysis

4️⃣ Ion Torrent Sequencing

It is the method to find the incorporation of a nucleotide by measuring the change in pH caused by the release of protons when a nucleotide is added. The change in PH is felt by a semiconductor chip in sequence data.

Applications:

❶ Whole genome sequencing
❷ Exome sequencing
❸ RNA sequencing
❹ Targeted sequencing
❺ Clinical diagnostics

5️⃣ Other Techniques

❶ Single Techniques
❷ Nanopore Sequencing            
❸ Helicose and Smart

4. The Key Benefits and Features

Whereas healthcare, research, and biotech sectors continue to thrive, next generation sequencing has taken center stage as a foundation technology. It also offers high throughput, scalability, and affordability.

Likewise, have a quick look at the key benefits and features of Next generation sequencing technologies. Those are,

4.1 Benefits:

👉 Massive Throughput

It may sequence the entire genome in a single attempt, also analyzing the millions of fragments concurrently. This makes it an ideal choice for a large-scale project such as population genetics, infectious disease surveillance, and cancer studies.

👉 Cost-Effective

The high throughput of NGS translates to lower costs per sample analyzed and makes large-scale genomic studies possible.

👉 Increased Sensitivity and Accuracy

The NGS can find rare variants and mutations with higher sensitivity and accuracy than existing methods, even at lower allele frequencies.

👉 Comprehensive Genome Analysis

However, Genetic testing may enable the analysis of entire genomes, transcriptomes, and epigenomes, providing a comprehensive view of the genetic information.

👉 Data Analysis

Altogether, it generates large amounts of data that require functional bioinformatics tools for analysis and interpretation.

4.2 Features:

Massively Similar Sequencing: This platform can sequence millions of DNA components, especially, enabling high-throughput breakdown.

Multifarious Sequencing Technologies: Moreover, there are a huge number of NGS technologies obtainable in the market, including Illumina sequencing and PacBio sequencing.

High-Quality Data: NGS platforms produce high-quality data with high resolution and accuracy and also support researchers to make meaningful discoveries.

Scalability: Eventually, these technologies are highly scalable and support researchers to analyze a wide range of test sample types and sequencing depths.

Rapid Turnaround Time: The NGS experiments can be completed quickly and allow researchers to get results within a minute.

Conclusion

In conclusion, the Next generation sequencing (NGS) is the future of genomics because it is rapid, precise, and cost-effective access to the analysis of DNA and RNA. NGS provides the capacity to sequence whole genomes within a much-reduced time frame, wasting it in old-fashioned counterparts such as Sanger sequencing.

Next generation sequencing is principally isolated with key steps such as DNA isolation, library preparation, sequencing, and analysis. Now, within the broad repertoire of Next Generation Sequencing techniques such as sequencing by synthesis, nanopore, and Ion Torrent can apply NGS from cancer research and molecular pathology to personalized medicine. The increasing NGS market phenomenon emerges with impacts shared by science and healthcare.