Next generation sequencing (NGS), also known as high-throughput sequencing, is an innovative DNA sequencing technology. It works on the concept of sequencing millions of DNA fragments in parallel at high speed and less cost. This has revolutionized the field of genomics. Let us explore the key developments and applications of NGS in detail.
Emergence of NGS Platforms
The first Next Generation Sequencing platform was developed by 454 Life Sciences in 2005 which could sequence up to 1 million DNA fragments at one time. Since then, different NGS platforms with advanced capabilities have emerged such as Illumina, Ion Torrent, SOLiD, PacBio, and Oxford Nanopore. All these platforms differ in sequencing chemistry, read length, speed, and accuracy. Illumina remains the most commonly used platform today due to its high throughput and accuracy.
Sequencing Chemistries Used
NGS works on different sequencing chemistries including sequencing by synthesis, sequencing by ligation, and real-time sequencing. Illumina uses sequencing by synthesis where fluorescent labeled nucleotides are detected as they are incorporated into DNA strands by DNA polymerase. Ion Torrent and Oxford Nanopore are based on real-time sequencing that detects hydrogen ions or changes in electrical current respectively when a nucleotide is added to DNA.
Applications of NGS in Genomics
NGS has rapidly advanced genomics research allowing sequencing of whole human genomes at a staggering pace and low cost. Some key applications include genome sequencing, transcriptome analysis,Chaperom e sequencing,detection of structural variations,epigenetic studies and more.
Genome Sequencing
NGS enables sequencing of entire genomes of organisms within a few hours or days. This has greatly facilitated research in human genetics, evolution, comparative genomics etc. Large scale genome projects like Human Genome Project and 1000 Genomes Project employed NGS.
Transcriptomics
RNA sequencing or transcriptomics is a powerful application of NGS to study whole transcriptomes. It provides quantitative measurement of gene and isoform expression levels and allows discovery of new transcripts, fusion genes and splicing variations.
Epigenomics
NGS enabled epigenomic profiling through techniques like ChIP-sequencing, Bisulfite sequencing to study DNA methylation, histone modifications and non coding RNAs at a global scale. This offers insights into epigenetic regulation of gene expression and diseases.
Single Cell Analysis
With advancement, NGS can now be applied for single cell analysis. This allows investigation of heterogenous cell populations and molecular profiling of rare cell types. Techniques like single-cell RNA sequencing and ATAC sequencing provide important functional information.
Clinical Applications
NGS finds widespread applications in clinical diagnostics and precision medicine. It is used for non-invasive prenatal testing,tumor genotyping,identification of disease causing mutations, microbiome analysis and pharmacogenomics. Examples include analyzing cancer genomes for better prognosis and therapies.
Challenges of NGS
While revolutionary, NGS also faces challenges like shorter read lengths, high capital investment for instruments, complex bioinformatics analysis of huge datasets,storage of sequencing data and need for well annotated reference genomes. Standardization of protocols and data analysis remain an active area of research.
Next generation sequencing emerged as a transformative technology that enabled large scale, low cost whole genome analysis. It has revolutionized genomics research and biomedical applications in a short span of time. Even though computational challenges exist, constant innovation is addressing issues to realize the full potential of NGS. Undoubtedly, NGS will continue to play a vital role in advancing our understanding of genomes and human health in the coming years.
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