Nanopore Sequencing Technology: The “Fourth-Generation Key” to Unlocking the Mysteries of Genes

In the field of gene sequencing, nanopore sequencing technology is sparking a technological revolution with its unique advantages. As the fourth-generation gene sequencing technology, it leverages the principles of single-molecule sequencing and innovative electrical signal detection, bringing unprecedented possibilities to life science research and clinical applications.

I. The Principle of Nanopore Sequencing: The Genetic Code in Electric Currents

The core of nanopore sequencing lies in combining single-molecule detection with electrical signal transmission. Unlike traditional sequencing methods, nanopore sequencing does not require complex PCR amplification or rely on optical signals. Instead, it directly reads DNA or RNA sequences through changes in electrical signals.

Specifically, nanopore sequencing utilizes a special nanopore protein that can be embedded into a high-resistance polymer film, forming an ionic channel. When a voltage is applied across the film, ions flow through the nanopore, generating an electrical current. The DNA or RNA molecules to be sequenced are then drawn through the nanopore in single-strand form by a motor protein. As different bases (A, T, C, G) pass through the nanopore, they cause varying degrees of current disruption. The sequencer records these fluctuations in electrical signals to deduce the order of the bases.

This electrical signal-based sequencing method not only simplifies sample processing but also significantly enhances sequencing efficiency. For example, nanopore sequencing can achieve a sequencing speed of up to 450 bases per second per pore. Additionally, nanopore sequencing boasts much longer read lengths compared to traditional technologies, with the longest reads reaching up to 4.2 Mb. This makes it highly advantageous for assembling complex genomes and discovering unknown structural variations.

II. The Advantages of Nanopore Sequencing: The Perfect Combination of Long Reads and Rapid Detection

Compared to traditional second-generation sequencing technologies (NGS), nanopore sequencing has made a leap from short reads to ultra-long reads. The advantage of long reads is their ability to more accurately span complex regions of the genome, such as highly repetitive sequences and structural variation regions, thereby improving the accuracy and completeness of sequencing.

Moreover, the rapid detection capability of nanopore sequencing also makes it shine in clinical applications. For example, in pathogen detection, nanopore sequencing can complete whole-genome sequencing of pathogens in a short time, quickly identifying the type of pathogen and its resistance genes, providing important evidence for clinical diagnosis and treatment.

III. Applications of Nanopore Sequencing: Broad Expansion from Laboratory to Clinical Settings

The application scope of nanopore sequencing is very broad, covering multiple fields including genomics, transcriptomics, epigenetics, and pathogen detection. In genomics research, it helps scientists more efficiently assemble complex genomes and reveal their structure and function. In pathogen detection, nanopore sequencing can rapidly identify infectious pathogens, including some that are difficult to detect with traditional methods.

Additionally, nanopore sequencing can directly detect modifications of DNA and RNA, such as methylation. This is of great significance for studying gene expression regulation and disease mechanisms.

With its unique principles and powerful functions, nanopore sequencing technology is becoming an indispensable tool in life science research and clinical diagnostics. As the technology continues to advance and its applications expand, nanopore sequencing will bring more surprises and breakthroughs to human health and life science research.

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