Sequencing technology platform and there are several common types ?
| Sequencing | Sequencing Platform | Sequencing Principle |
|---|---|---|
| Generation Sequencing | ABI3730, ABI3500 | Dideoxy termination method, capillary electrophoresis |
| Generation Sequencing | GS 454、SoLiD、Illumina HiSeq | Pyrosequencing, sequencing connected sequencing by- synthesis method |
| Three generations of sequencing | PacBio RS, Oxford In-opore Min-lon | Single molecule sequencing, nanopore sequencing |
Sequencing technology has so far undergone three generations of development.
In fact, as early as after the DNA double helix structure found soon, called degradation method (sequencing by degradation, SbD) DNA sequencing technology would have been invented, but the method is complicated to operate, erratic results, at the time not to promote open carried out large-scale application. Then, Sanger and Coulson, 1975 invented the "Plus and Minus" DNA sequence was determined (Sangerand Coulson, 1975); Maxam and Gilbert , 1977 invented chemical degradation sequencing. Finally in the same year, the Sanger incorporated ddNTPs (dideoxynucleoside triphosphates) in the nucleic acid polymerization reaction, well-known dideoxy chain termination method of birth (Sanger, Nicklen, and Coulson, 1977). In addition, in the same period, other sequencing methods such as ligase sequencing method (sequencing by ligation, SbL), pyrosequencing (Pyrosequencing), sequencing by hybridization method (sequencing by hybridization, SbH) ( Drmanac R.et al, 2001.; Drmanac R.et al, 1989; Drmanac S.et al, 1998) and the like, due to technical limitations not get large-scale application, these techniques are collectively referred to. the first generation sequencing .
The second generation sequencing technology is accompanied by the sequencing of the birth of the Human Genome Project (Schuster, 2008). Since 1990, with the increasing scale sequencing, high-throughput sequencing platform, a second-generation sequencing technologies have emerged (Fuller ef al, 2009;. Mardis, 2008; Shendureand Ji, 2008; Wang Xi, etc., 2010). Generation sequencing there are three sequencing method: Ilumina's by- synthesis techniques , developed in the art on the basis of a Genome Analyzer sequencing platform; Roche's pyrosequencing , sequencing 454 also known, in which Based on the technology, Roche has developed GSFLX, GSJunior sequencing platforms; ABI company uses technology developed connection Solid sequencing platform.
Recently, third-generation sequencing technology has been developed to PacBio's SMRT and Oxford Nanopore Technologies company nanopore single-molecule sequencing technology as the representative (Klumpp, Fouts, and Sozhamannan, 2012; Niedringhaus et al, 2011.). Wherein the basic principle PacBio SMRT: DNA template and polymerase binding, 4 color fluorescent labeled four bases (dNTPs), different pairs of substantially different color fluorescence. While the DNA polymerase is one key to achieving long read length. While this technique sequencing is fast, but the sequencing error rate, reaching 15%. Error correction can only be carried out by multiple sequencing. Oxford Nanopore Technologies's single nanopore single-molecule sequencing technology is different from the conventional sequencing technology, which is based on an electrical signal sequencing techniques. The key to this technique is to design a special nanopore, when single-strand DNA bases through the nanopore, current intensity flowing through the nanopore will change. From the change in the current to identify a different base, for the purpose of sequencing.
