Next-Generation Sequencing (NGS)

What is Next-Generation Sequencing (NGS)?

Next-generation sequencing (NGS) describes several techniques used to determine the sequence of DNA or RNA. Scientists can use that sequencing data to study genetic variations or mutations associated with diseases or biological conditions. Because NGS allows scientists and researchers to sequence thousands or even millions of DNA molecules simultaneously, it has significantly reduced the cost and the amount of time needed to sequence genes. That has led to dramatic advances in clinical diagnostics, genetic diseases, and personalized medicine.

Sanger Sequencing vs. Next-Generation Sequencing

Sanger sequencing and next-generation sequencing (NGS) are similar in concept, but they have profound differences. Both next gen sequencing and Sanger sequencing involve using DNA polymerase to add fluorescent nucleotides one at a time onto a growing DNA or RNA template strand. The fluorescent tags identify each incorporated nucleotide.

Where these two RNA and DNA sequencing methods differ is in each method’s sequencing volume. The Sanger method sequences DNA or RNA one strand at a time, while next-generation sequencing – initially called “massively-parallel sequencing” – can simultaneously sequence hundreds to thousands of genes at one time.

Sanger sequencing reagents produce exceptional results, and Sanger sequencing is still considered the “gold standard” for clinical research sequencing with a 99.99% accuracy rate. However, NGS technology in clinical research labs is expanding as labs benefit from their higher throughput and cost-efficiency.

Advantages of NGS

Next-generation sequencing provides multiple benefits when compared to Sanger sequencing, including:

• Can sequence an entire genome at the same time
• Faster turnaround time
• More cost-effective
• Requires lower sample input
• Improved accuracy and reliability
• Higher sensitivity allows detection of variants at lower frequencies

Typical NGS Workflow

Whether you are conducting DNA sequencing or RNA sequencing, your workflow will follow the same basic steps:

1. Library preparation includes fragmenting DNA or RNA from the rest of the sample, preparing DNA or RNA for sequencing by adding adapters and checking all fragments for size and quality before adding them to the sequencing library
2. Clonal amplification generates multiple copies of target areas of interest within a DNA or RNA sample to ensure they provide a robust and detectable signal during the sequencing process.
3. Sequencing is the process that tells scientists what genetic information each DNA or RNA molecule contains.
4. Data analysis is the process of applying a wide range of analytical methods to DNA or RNA sequencing information to determine its structure, function, features, or other characteristics.

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