Gene sequencing "Moore's Law" first appeared: three generations of sequencing to the "second generation" life?
Recently, Xingzheng Medicine Health released a report on the introduction of the three-generation gene sequencing industry. The website believes that this report will help medical practitioners have a comprehensive understanding of the three-generation sequencing technology and market prospects, and thus reprinted and sort out.
In the case that “Second Generation Sequencing†(NGS) has not yet ushered in an investment boom, the “three-generation sequencing†(3GS), which has repeatedly broken through the technology, has entered the field of investors. In 1986, the first commercial gene sequencing equipment was officially launched, and the second generation of sequencing equipment appeared, with a period of 19 years. The second generation of equipment came out, and the birth of the third generation of equipment took only five years, and the speed of updating the genetic sequencing equipment is accelerating. This is like the "2G" mobile phone skipping "3G" and directly crossing the "4G" era.
The report analyzes third-generation genetic testing techniques in four ways:
1. The development status of the third generation genetic testing technology
The three-generation sequencing technology represented by Helicos' Heliscope single-molecule sequencer, Pacific Biosciences' SMRT technology and Oxford Nanopore Technologies' nanopore single-molecule technology has gradually matured after years of development.
Although the current technology has the problems of high cost, high error rate and insufficient bioinformatics analysis software, it has obvious advantages in terms of reading length and sequencing speed.
The third-generation sequencing equipment has achieved stability and miniaturization. In the future, with the improvement of accuracy, parallel sequencing capability and enzyme activity, the third-generation sequencing technology will become an important technological trend in the future development, and large-scale commercialization will be The trend of the times.
2. Principles of the third generation gene sequencing method
Helicos's Heliscope single-molecule sequencer, Pacific Biosciences' SMRT technology and Oxford Nanopore Technologies' nanopore single-molecule technology are considered third-generation sequencing technologies.
Compared with the previous two generations of technology, their biggest feature is single-molecule sequencing. Among them, Heliscope technology and SMRT technology use fluorescent signals for sequencing, while nanopore single-molecule sequencing technology uses electrical signals generated by different bases for sequencing.
PacBio SMRT technology uses the idea of ​​sequencing while synthesizing, and uses SMRT chip as the sequencing carrier. There are many small holes on the chip, and there are DNA polymerases in each well.
The basic principle of sequencing is: DNA polymerase and template are combined, and 4 colors are fluorescently labeled with 4 bases (ie, dNTP). In the base pairing stage, different bases are added, and different light is emitted, depending on the wavelength and peak of the light. Determine the type of base entered. DNA polymerase is one of the keys to achieving ultra-long read length. The read length is mainly related to the activity of the enzyme, which is mainly affected by the damage caused by the laser.
In addition, some base modifications can be detected by detecting the sequencing time between two adjacent bases. If the base is modified, the speed at the time of passing through the polymerase will be slowed down, and the distance between adjacent two peaks. Increase, you can use this to detect information such as methylation. The sequencing of SMRT technology is fast, with a few dNTPs per second.
However, at the same time, its sequencing error rate is relatively high (this is almost a common problem with single-molecule sequencing technology), reaching 15%. Fortunately, its error is random, and there is no sequencing error bias like the second-generation sequencing technology, so it can be effectively corrected by multiple sequencing (at the cost of repeated sequencing, that is, the cost will increase) .
SMRT technology schematic
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