ABSTRACT
A number of NGS platforms for RNA-seq and other applications have been developed. Several major NGS platforms are briefed here. Illumina’s platform (https://www.illumina.com/) represents one of the most popularly used sequencing by synthesis chemistry in a massively parallel arrangement. Currently, it markets HiSeq X Five and HiSeq X Ten instruments with population power; HiSeq 2500, HiSeq 3000, and HiSeq 4000 instruments with production power; Nextseq 500 with exible power; and MiSeq with focused power. e HiSeq X Ten is a set of 10 ultra-high-throughput sequencers, purpose-built for large-scale human whole-genome sequencing at a cost of $1000 per genome, which together can sequence over 18,000 genomes per year. e MiSeq desktop sequencer allows you to access more focused applications such as targeted gene sequencing, metagenomics, small-genome sequencing, targeted gene expression, amplicon sequencing, and HLA typing. New MiSeq reagents enable up to 15 GB of output with 25 M sequencing reads and 2 × 300 bp read lengths. Life Technologies (https://www.lifetechnologies. com/) markets sequencing by oligonucleotide ligation and detection (SOLID) 5500 W Series Genetic Analyzers, Ion Proton™ System, and the Ion Torrent™ Personal Genome Machine® (Ion PGM™) System. e newest 5500 W instrument uses ow chips, instead of beads, to amplify templates, thus simplifying the workow and reducing costs. Its sequencing accuracy can be up to 99.99%. e Ion Proton™ and Ion PGM™ Systems
are ion semiconductor-based platforms. e Ion PGM™ System is one of top selling benchtop NGS solutions. Roche markets 454 NGS platforms (https://www.454.com/), the GS FLX+ System, and the GS Junior Plus System. ey are based on sequencing by synthesis chemistry. e GS FLX+ System features the unique combination of long reads (up to 1000 bp), exceptional accuracy and high-throughput, making the system well suited for larger genomic projects. e GS Junior Plus System is a benchtop NGS platform suitable for individual lab NGS needs. Pacic Biosciences (https://www.pacicbiosciences.com/) markets the PACBIO RSII platform. It is considered as the third-generation sequencing platform since it only requires a single molecule and reads the added nucleotides in real time. e chemistry has been termed SMRT for single-molecule real time. e PacBio RS II sequencing provides average read lengths in excess of >10 KB with ultra-long reads >40 KB. e long reads are characterized by high 99.999% consensus accuracy and are ideal for de novo assembly, targeted sequencing applications, scaolding, and spanning structural rearrangements. Oxford Nanopore Technologies (https:// nanoporetech.com/) markets the GridION™ system, the PromethION, and the MinION™ devices. Nanopore sequencing is a third-generation single-molecule technique. e GridION™ system is a benchtop instrument and an electronics-based platform. is enables multiple nanopores to be measured simultaneously and data to be sensed, processed, and analyzed in real time. e PromethION is a tablet-sized benchtop instrument designed to run a small number of samples. e MinION device is a miniaturized single-molecule analysis system, designed for single use and to work through the USB port of a laptop or desktop computer. With continuous improvements and renements, nanoporebased sequencing technology may gain its market share in not distant future.
