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The alternation of hunter and gatherer roles: the evolutionary truth behind the sexual division of labor?
In the world of genomics, the evolution of sequencing technology has had a revolutionary impact on our understanding of the composition and operation of life. Among them, single-molecule real-time (SMRT) sequencing, as an innovative technology, is reshaping our understanding of the genome with its unique operating principles and efficient performance. The core of SMRT sequencing lies in zero-mode waveguide (ZMW) technology, which allows researchers to track the extension process of DNA polymerase at the single-molecule level and ultimately decode genomic messages.
SMRT sequencing can sequence DNA at the single-molecule level, allowing us to obtain more accurate genomic information.
Technical Basics
SMRT sequencing relies on a superior optical technology. In each ZMW, a single DNA polymerase is immobilized to the bottom, and four different fluorescent dye-labeled nucleotides are added one at a time as the DNA template extends. When the polymerase data captures a fluorescent signal, the corresponding nucleotide is determined. This process is the key to real-time sequencing.
The use of fluorescent dyes allows the detector to accurately identify the addition of each nucleotide during polymerase synthesis.
Sample preparation and sequencing performance
Prior to sequencing, DNA fragments are converted into circular structures during the library preparation stage. This approach allows us to read the same molecule multiple times, significantly increasing read length and accuracy. According to the report, using the latest Sequel 6.0 chemistry, the average read length of shorter insert libraries can reach 100,000 base sequences, while the average read length of longer insert libraries is approximately 30,000 bases.
The launch of Sequel 6.0 chemical technology has greatly improved sequencing accuracy and data output, which is particularly important for genomic research.
Historical development
PacBio commercialized SMRT sequencing technology in 2011 and subsequently introduced RS and RS II model instruments, which have been improved over time to improve sequencing accuracy and read length. In 2013, PacBio launched new DNA polymerase binding reagents that not only increased read length but also improved variant identification.
Application scope
SMRT sequencing is used in many areas, including exploring novel genomes, identifying genetic variants, and even studying complete genomes of bacteria and archaea. Its ultra-long read length can help scientists piece together genomes more accurately and effectively identify different transcript variants of genes.
Ultra-long read technology allows researchers to capture the entire genome without any hindrance, opening a new chapter in genomics research.
With the increasing maturity of genomics technology, SMRT sequencing not only provides us with tools to explore the mysteries of life, but also triggers in-depth thinking about the nature of life. How will this technology further advance the development of life sciences and uncover the unknown, and how will our understanding of all this evolve?
