Differences between different generations of high-throughput sequencing technology (NGS)

Different generations of high-throughput sequencing (NGS) technologies have significant differences in many aspects, including sequencing principles, throughput, cost, accuracy, read length, and application fields. Below is a detailed comparison of first-, second- and third-generation high-throughput sequencing technologies:

1. First-generation sequencing technology

1>Features:

Sequencing read length: The read length of first-generation sequencing technology is longer, usually up to 1000bp or even longer.

Accuracy: The accuracy is extremely high, reaching 99.999%.

Throughput and cost: However, its sequencing cost is high and its throughput is low, which limits its application in large-scale projects.

2>Application: Despite the problems of throughput and cost, first-generation sequencing technology still plays an important role in research requiring highly accurate sequence information due to its high accuracy.

2. Next-generation sequencing technology (NGS)

1>Features:

Parallel sequencing: Using a parallel sequencing strategy, millions or even billions of DNA molecules can be sequenced simultaneously, greatly improving sequencing throughput.

Cost and speed: The cost of sequencing has been significantly reduced, and the sequencing speed has been significantly improved. It used to take years to sequence a human genome, but using next-generation sequencing technology it can be completed in weeks or even days.

Accuracy: Maintains high accuracy, but has shorter read lengths compared to first-generation sequencing technology.

2>Application:

Genomics and transcriptomics: widely used in whole genome sequencing, transcriptome sequencing and other fields.

Clinical testing: plays an important role in genetic disease screening, prenatal testing, tumor diagnosis, etc.

3. Third-generation sequencing technology

1>Features:

Single-molecule sequencing: Compared with the first two generations of technology, the biggest feature of the third-generation sequencing technology is single-molecule sequencing, which does not require PCR amplification and can directly sequence a single DNA or RNA molecule.

Long read length: The read length has increased significantly, reaching the Mb level, far exceeding the second-generation sequencing technology.

Real-time sequencing: Sequence information can be obtained in real time to meet the needs of dynamic detection.

Low cost and portability: The cost of sequencing equipment has been reduced, and some equipment is portable, making it convenient for on-site sequencing.

2>Application:

Large genome splicing: In genome splicing, the completeness and accuracy of the genome can be significantly improved due to the characteristics of long reads.

Full-length transcriptome analysis: RNA can be directly sequenced to accurately identify multiple homologous isomers of each gene without interruption and reverse transcription.

Structural variation detection: The ability to accurately detect large fragments of structural variation, such as deletions, inversions, and translocations, is of great significance to disease research.

Rapid identification of microorganisms: Due to its real-time and rapid characteristics, sequencing can be performed directly at the collection point to quickly complete species classification and identification.

There are significant differences in many aspects between different generations of high-throughput sequencing technologies. These differences make different prediction technologies suitable for different application fields. In the future, high-throughput prediction technologies will continue to improve and expand their application fields, providing Provide strong support for human health and life science research.