Next-Generation Sequencing and Emerging Technologies

Abstract

Genetic sequencing technologies are evolving at a rapid pace with major implications for research and clinical practice. In this review, the authors provide an updated overview of next-generation sequencing (NGS) and emerging methodologies. NGS has tremendously improved sequencing output while being more time and cost-efficient in comparison to Sanger sequencing. The authors describe short-read sequencing approaches, such as sequencing by synthesis, ion semiconductor sequencing, and nanoball sequencing. Third-generation long-read sequencing now promises to overcome many of the limitations of short-read sequencing, such as the ability to reliably resolve repeat sequences and large genomic rearrangements. By combining complementary methods with massively parallel DNA sequencing, a greater insight into the biological context of disease mechanisms is now possible. Emerging methodologies, such as advances in nanopore technology, in situ nucleic acid sequencing, and microscopy-based sequencing, will continue the rapid evolution of this area. These new technologies hold many potential applications for hematological disorders, with the promise of precision and personalized medical care in the future.

Genetic Background of von Willebrand Disease: History, Current State, and Future Perspectives

“…Alternatively, direct screening of arginine hotspot regions of VWF (eight CGA codons), which may be converted into a stop codon; screening of the most mutated exon 28 alone; sequencing of the discrete regions of the VWF gene (in type 2 patients) using traditional Sanger sequencing method. 74 The Sanger method allows establishing the nucleotide sequence of a DNA fragment (direct DNA sequence analysis). This technique is the most frequently adopted for the identification of VWF mutations.…”

Whole genome sequencing for the genetic diagnosis of heterogenous dystonia phenotypes

“…Whole genome sequencing (WGS) offers several advantages over other targeted genetic sequencing approaches [8]. By querying the entire genome, it offers both uniform coverage of coding and noncoding regions of the genome and allows for accurate detection of copy number variants (CNVs) and structural variants (SV).…”

Think before you prescribe: how dentistry contributes to antibiotic resistance

“…A read is a section of a unique fragment of nucleic acid. NGS platforms are based on different methods of sequencing, and include sequencing by synthesis (Illumina/Solexa), single‐molecule real‐time sequencing (Pacific Biosciences), nanopore (Oxford Nanopore) and ion semiconductor (Ion Torrent) . The platforms are compared by read size (short or long), number of reads that can be sequenced per library and the total amount of data generated …”

“…NGS platforms are based on different methods of sequencing, and include sequencing by synthesis (Illumina/Solexa), single-molecule real-time sequencing (Pacific Biosciences), nanopore (Oxford Nanopore) and ion semiconductor (Ion Torrent). 25 The platforms are compared by read size (short or long), number of reads that can be sequenced per library and the total amount of data generated. 25 Metagenomic approaches aim to analyse all or as much of the genetic material which can be recovered directly from a specific sample, preserving the microbial complexity and biodiversity of the niche environment so that it can be studied as a whole.…”

“…25 The platforms are compared by read size (short or long), number of reads that can be sequenced per library and the total amount of data generated. 25 Metagenomic approaches aim to analyse all or as much of the genetic material which can be recovered directly from a specific sample, preserving the microbial complexity and biodiversity of the niche environment so that it can be studied as a whole. Culturedependent studies of oral bacteria (limited to species that can be cultivated in vitro) are virtually redundant, as NGS studies have shown that approximately 68% of oral bacteria cannot be cultured.…”