The difference between targeted sequencing and high-throughput sequencing

With the rapid development of genomics and molecular biology, gene sequencing technology has become an important tool in many fields such as medicine, agriculture, and environmental protection. Among many sequencing technologies, targeted sequencing and high-throughput sequencing are often mentioned, but what are the differences between the two? The editor of Hechuang will introduce in detail the difference between targeted sequencing and high-throughput sequencing, and help you better understand their applications.

1. Basic concepts 1. Targeted sequencing, as the name suggests, refers to only focusing on the target region or specific gene sequence when performing genome sequencing. Targeted sequencing uses specific probes or primers to enrich only the region of interest, thereby improving the sensitivity and accuracy of sequencing. This approach can help researchers focus more on analyzing mutations or mutations in specific genes. 2. High-throughput Sequencing (HTS) is a technology that can sequence a large number of DNA molecules at the same time. Its characteristic is that it has extremely high parallel processing capabilities and can obtain massive genomic data at one time. Compared with traditional Sanger sequencing, high-throughput sequencing not only greatly improves the sequencing speed, but also significantly reduces the sequencing cost and is widely used in genomic research, clinical testing and other fields.

2. Differences in technical principles 1. Targeted sequencing principle The core of targeted sequencing lies in "targeting". Typically, researchers first select one or more gene regions of interest that may be associated with the disease or have special biological significance. These regions are captured by using specific probes or primers, and the captured DNA fragments are sequenced. This method is suitable for situations where high-precision analysis is required, and can accurately find variation information in the target area. 2. High-throughput sequencing principle. High-throughput sequencing is to sequence a large number of DNA samples through parallelization. The core of its technology lies in cutting DNA samples into small fragments, and splicing and aligning these fragments through complex algorithms to finally obtain a complete genomic sequence. High-throughput sequencing is not only suitable for whole genome sequencing (WGS), but can also be used in a variety of applications such as transcriptome sequencing (RNA-Seq) and methylation sequencing. The advantage is that it can obtain massive data, but the sequencing depth of the target region may not be as good as targeted sequencing.

3. Different application scenarios 1. Targeted sequencing is mainly used to conduct detailed analysis of specific genes, mutations or mutations. For example, in cancer research, targeted sequencing can help researchers focus on known cancer-related genes and accurately identify gene mutations related to tumors, thereby providing a basis for personalized treatment. In clinical diagnosis, targeted sequencing can be used to detect some specific genes related to genetic and infectious diseases. 2. The application of high-throughput sequencing is more extensive. In addition to being used for whole genome sequencing, it can also support various genomic research, such as gene expression, variant detection, microbiome analysis, etc. High-throughput sequencing is suitable for research scenarios that require a large amount of data, such as in Genome-wide Association Studies (GWAS), which can provide a large amount of sample data support, helping scientists find genetic mutations related to diseases.

4. The difference between sequencing depth and data volume 1. A significant feature of targeted sequencing is that it can obtain higher sequencing depth in specific target areas. Since only focusing on specific genes or regions, targeted sequencing can provide more accurate variant detection results than high-throughput sequencing. Therefore, it is particularly prominent in the detection of low-frequency variations, and is especially suitable for accurate analysis of rare mutations. 2. In contrast, the amount of data for high-throughput sequencing is very large, and it can generate genome-wide data at one time. Nevertheless, since its sequencing depth is not as high as targeted sequencing, it may be insufficient when dealing with rare variants in the genome. However, the advantages of high-throughput sequencing lies in its wide application and processing capabilities of massive data, which are suitable for large-scale genomic analysis.

5. Differences in cost and time From a cost perspective, targeted sequencing is usually cheaper than high-throughput sequencing. Since targeted sequencing only focuses on gene sequences in specific regions, it requires a small sample size and a low data processing volume, so it is cost-effective. While high-throughput sequencing can process more samples and more data, it is relatively expensive because it involves the entire genome or large amounts of genetic data. In terms of time, targeted sequencing usually takes less time than high-throughput sequencing. The process of targeted sequencing is relatively concentrated and focuses on specific regions, so its workload is relatively small. High-throughput sequencing requires more time to analyze and process data, especially when whole genome sequencing, the huge amount of data may take longer to complete.

Targeted sequencing and high-throughput sequencing have obvious differences in technical principles, application scope, sequencing depth, data volume and cost. Targeted sequencing is suitable for accurate analysis of specific regions or genes, while high-throughput sequencing is suitable for large-scale genomic data analysis. Both have their own advantages and disadvantages and play an important role in different research and clinical applications.

Related reading recommendations

Pipette White Paper: Magnetic Modules

Overview of pipetting robot instrument services

Introduction to the temperature control module of Opentrons pipette

Automation PCR e-book

Opentrons Flex Transfer Handle

Why is it necessary to automate nucleic acid preparation