Definition of NGS
Next-generation sequencing (NGS), also known as high-throughput sequencing, is a method of analyzing genetic material that allows for the rapid sequencing of large amounts of DNA or RNA. In contrast to traditional sequencing techniques, NGS can simultaneously sequence hundreds to thousands of genes or whole genomes in a short period of time, allowing researchers to expand the scale and discovery bandwidth of their studies.
The entire workflow of NGS, from sample source to assessed data, can be broken down into 4 Steps:
Overview of the NGS Workflow
Nucleic acid extraction is a standard upstream necessity of the NGS workflow. To be suitable for NGS, DNA and RNA must be pure and free of contaminants such as salts, proteins, and phenol. Zymo Research’s comprehensive portfolio of DNA extraction and RNA extraction kits ensure that inhibitor-free, highly concentrated nucleic acid can be accurately analyzed using NGS. Following purification, the DNA and/or RNA must then be prepared into a format that can be read by a sequencing instrument. The purified DNA and/or RNA cannot be read “as is” following extraction by the sequencer. DNA and/or RNA libraries must be prepared using the following steps to be identified by the sequencing instrument.
- Fragmentation: Converts large, intact genetic material into smaller insert sizes for short-read sequencing technology utilized by Illumina.
- Adapterization: Special adapters, or short DNA sequences, are added to the ends of the nucleic acid fragments, which allow them to be recognized and attached to the sequencing platform for amplification during the sequencing reaction.
The methodology of fragmentation and adapterization is what distinguishes one library preparation protocol from another. Additional steps are included in the library preparation procedure that allow for specific desired analyses as well as for pooling multiple samples into one sequencing run.
Key Features of NGS
- Ultra-High-Throughput - Massively Parallel Sequencing (MSP), the technology used for NGS applications, is capable of sequencing millions of DNA or RNA fragments at the same time, generating large amounts of molecular data.
- High Accuracy & Precision: NGS enables single base pair resolution of genetic material.
- Scalable: This technology allows researchers to read millions of DNA or RNA fragments simultaneously, making large-scale sequencing accessible and practical.
- Faster Sequencing Rate: An entire genome can be sequenced in a single day using NGS technology.
Applications of NGS
NGS has become an everyday tool to address a wide array of questions scientists ask. Some of the most common applications and fields requiring NGS are listed below.
- Cancer Research: Sequencing samples from cancer patients helps clinicians detect rare somatic variants, tumor subclones, and circulating DNA fragments. NGS can be used to diagnose cancer in early stages using liquid biopsy, as well as inform treatment options.
- Complex Disease Research: Scientists can use NGS to better understand the molecular underpinnings of autoimmune and rheumatic diseases, atherosclerosis, neurological diseases, and psychiatric disorders.
- Reproductive Health: Sequencing and array technologies provide information on fetal anomalies to guide reproductive health choices and improve prenatal care.
- Drug Discovery: NGS can help identify targets for pharmaceutical agents, which can lead to the development of more effective drugs.
- Microbiology: Sequencing the genomes of microbes, bacteria, and viruses can help identify pathogens, track outbreaks, diagnose infectious diseases, and study antimicrobial resistance.
- Forensic Genomics: NGS technology provides more accurate, comprehensive insights from forensic DNA than traditional methods, resulting in more conclusive identification results.
A Comprehensive Portfolio of NGS Preparation Methods
NGS Library Preparation methods offered by Zymo Research utilize various approaches to prepare nucleic acids for sequencing. The portfolio of solutions offered has been optimized with virtually no limit on the sample source from which the nucleic acids are extracted, or the species. Understanding that every sequencing project is unique, requiring a tailored approach to work within the limitations of the sample, Zymo has provided solutions of products and NGS services. Whether it’s low input, compromised nucleic acids integrity, or unique sample sources, the Zymo-Seq portfolio helps leveraging the power of Next-Generation Sequencing to propel the researcher’s project forward.