a brief introduction to next generation sequencing
by Neoteryx | 1 min read
NGS sequencing includes Illumina, Roche 454, Ion Torrent, and Solid sequencing.
It’s a single stroke sequencing of vast numbers of short threads. Illumina sequencing utilizes approximately100-150bp of reads. On the other hand, you should tie up longer fragments up to generic adaptors, then hardened to a slide using adaptors. Moreover, amplifying the reads using Polymerase chain reaction (PCR) individually creates a spot with numerous copies of the same read. Separation of each read into single strand is then done before sequencing.
First of all, like Illumina, it uses longer reads. It does this by reading optical signals by sequencing multiple reads at once as you add their bases. Secondly, you should split RNA/DNA into 1kb of shorter reads. Furthermore, Polymerase chain reaction (PCR) amplifies the fragments using specific primers and placed in a single well of a slide. Finally, flooding of the slide with one of the four nucleoside triphosphate (NTP) is then done for sequencing.
ION Torrent/Proton Sequencing
ION Torrent/Proton doesn’t make use of optical signals like Illumina and 454 sequencing. They capitalize on the fact that the addition of dNTP to DNA polymer releases H+ ion. However, like other NGS you should split input RNA/DNA by ~200bp.
Applications of Mitra Microsampling Technology in NGS
The Mitra microsampler allows the production of accurate results from molecular research labs. Blood collection via Mitra microsampler is credited for being non-evasive, less costly, and very efficient. Secondly, the immediate benefits provided are working with volumetric accurate dried blood spot which reduces the cost of incidental reworks. Moreover, it’s amenable to RNA/DNA isolation and purification using central labs kits. The blood collection is convenient to the donor, and comes without expensive refrigeration or special transportation. Finally, the samples are compatible with Next Generation Sequencing platforms.
The main advantages of NGS over traditional Sanger sequencing include speed, cost, accuracy, and the sample size. In addition, NGS requires less RNA/DNA than Sanger sequencing.