Subsequent-generation sequencing (NGS), generally known as “deep sequencing” or “massively parallel sequencing,” is a DNA sequencing approach that has paved the manner for main development in medical and translational analysis. Clinicians and researchers use the approach to check the underlying DNA mechanisms related to neonatal and infectious ailments, uncommon genetic issues, cancers, and extra. Research into these ailments and issues have laid the groundwork for the improvement of better-targeted, customized therapies.
Though next-generation sequencing provides loads of advantages, each in the clinic and in analysis, it doesn’t come with out its disadvantages. The life sciences journal BioTechniques is an effective place to learn up on these execs and cons. The journal is well known for overlaying the efficacy and reproducibility of methods and strategies in science and drugs, fairly than focusing wholly on therapies.
Right here, we’ll work by means of the phases of NGS and focus on its execs and cons and its position in a range of functions.
The Phases of Next-Generation Sequencing
The NGS workflow includes 4 essential phases. The primary is pattern preparation, throughout which a researcher extracts genomic DNA from a pattern. The pattern is normally saliva, blood, or tissue. The researcher fragments the DNA into shorter sequences and follows up with ligation of adapters, amplification, and enrichment.
The following stage is library preparation. Throughout this stage, the researcher randomly fragments the DNA or cDNA. They normally obtain this by means of sonication or by making use of an enzymatic remedy. The platform they use dictates the optimum fragment size.
The third stage is the sequencing stage. The researcher chooses a sequencing methodology based mostly on their platform. Some instance strategies embody pyrosequencing, sequencing by synthesis or ligation, and reversible terminator sequencing. Sequencing by synthesis is one of the mostly chosen strategies as this strategy permits researchers to sequence heaps of DNA directly and at excessive sensitivity. With this strategy, researchers can detect a wide variety of genetic alterations, resembling structural variants, small insertions and deletions, and single-nucleotide polymorphisms (SNPs).
The ultimate stage of the workflow is knowledge evaluation. The researcher makes use of knowledge evaluation functions or bioinformatic instruments to pinpoint pathogenic variants, align to the reference sequence, and carry out high quality management checks.
Pros of Next-Generation Sequencing
Earlier than scientists developed NGS, they carried out Sanger (first-generation) sequencing, which was prevalent for 30 years and shaped the foundation of scientists’ understanding of the human genome. Now, NGS has outmoded Sanger sequencing as a result of it provides a extra environment friendly workflow, improved sensitivity, and protection. It’s additionally far more cost-effective: The price of sequencing the human genome has fallen over the previous decade from $20-$25 million to underneath $1,000 by 2016.
The time required to hold out NGS and obtain outcomes has additionally dropped over current years. In the present day, NGS platforms can sequence hundreds of thousands of DNA fragments concurrently, which means that researchers can sequence nearly something inside a day, from particular goal areas to the whole human genome. It takes roughly 10 days to obtain a whole-genome sequencing report from the day the lab receives the tumor specimen.
On high of this, NGS makes it attainable for researchers to determine abnormalities throughout the complete genome. This implies they’ll sequence abnormalities throughout insertions, deletions, substitutions, duplications, chromosome inversions/translocations, and copy quantity adjustments (gene and exon). NGS may determine abnormalities throughout the whole genome utilizing much less DNA than the DNA required for conventional sequencing strategies.
Cons of Next-Generation Sequencing
Regardless of all the advantages of NGS, the approach does have some cons. First, though NGS gives data on many molecular aberrations, the medical significance of many recognized abnormalities remains to be unknown.
Second, NGS requires massive knowledge storage capabilities, refined bioinformatics programs, and quick knowledge processing infrastructures, every of which will be pricey. Though some establishments have the funding to fulfill these necessities, others can’t fund the workers or computational assets to interpret and analyze such excessive knowledge masses.
Third, whereas researchers can use NGS to sequence a complete DNA sequence, they’ll solely use knowledge from roughly 3% of the genome in medical follow. So, NGS has far more potential in the analysis area than it does in the medical area.
Next-Generation Sequencing Purposes
Regardless of NGS’ limitations, the approach has turn out to be integral to a lot medical and scientific analysis. Scientists have employed NGS in a number of analysis functions, resembling whole-genome sequencing to determine an organism’s full DNA sequence, whole-exome sequencing to research a genome’s coding areas, focused sequencing to look at particular genomic areas, epigenomics to evaluate epigenetic modifications, and PCR for next-generation polymerases in the NGS workflow. Researchers have additionally used NGS in RNA sequencing to carry out transcriptome profiling of coding and non-coding areas, determine genes in particular cell varieties, and decide genetic alterations resembling gene fusions and single nucleotide variants (SNVs).
The newest developments in NGS have even seen the approach advance therapies that improve COVID-19 testing and fight weight problems. As scientists proceed to form NGS applied sciences, sequencing methods ought to turn out to be much more cost-effective and accessible.
Publishing Advances in Life Science Applied sciences
BioTechniques has been reviewing laboratory strategies and methods since 1983. Since then, the journal has grown a worldwide viewers of scientists and analysis professionals who concentrate on fields spanning from the life sciences, chemistry, and physics to pc science, plant science, and agricultural science. Not solely do these customers profit from the print journal, however in addition they discover a wealth of assets on BioTechniques’ multimedia web site. These assets embody articles, eBooks, movies, interviews, webinars, and podcasts, which delve into laboratory strategies like next-generation sequencing, western blotting, polymerase chain response, chromatography, and CRISPR gene enhancing.
BioTechniques is one of the 34 peer-reviewed, open-access journals that Future Science Group publishes. The progressive medical and scientific writer can also be residence to titles like Regenerative Drugs, Nanomedicine, and Future Oncology. Future Science Group receives over 5 million article downloads yearly and has printed over 50,000 articles to date.
Attention-grabbing associated article:
