Summary of this article
The MiSeq is a next-generation sequencer (NGS) from Illumina, Inc. that is primarily suited for targeted sequencing, small-scale whole genome analysis, gene expression analysis, and microbial genome analysis (NovaSeq), and is positioned as the model of choice for small and medium-scale sequencing analyses in laboratories and clinical laboratories.
1. main features of MiSeq
- Compact design:
- MiSeq is smaller than larger sequencers (HiSeq and NovaSeq) and is suitable for use in the laboratory and clinical settings. It is also designed in an easy-to-handle size, providing a readily available environment for next-generation sequencing.
- Rapid data acquisition:
- A key feature of MiSeq is its ability to obtain sequencing results in a short period of time. Even for full-run analysis, data can be obtained within one day, making it suitable for projects that require quick results.
- High flexibility:
- MiSeq can accommodate a variety of sequencing projects from short to long read lengths. Single-ended or paired-end reads can be selected, and read lengths up to 300 base pairs (bp) are supported.
- Data Throughput:
- MiSeq can generate up to 15 Gb of data, giving you the flexibility to adjust the sequencing depth according to your research objectives. It can handle sequencing of small target regions as well as mid-size genome analysis.
- Sequencing Bisynthesis (SBS) technology:
- Like all Illumina sequencers, the MiSeq uses Sequencing by Synthesis (SBS) technology, which reads bases one by one through a DNA synthesis reaction, providing highly accurate data.
2. the MiSeq technology
Sequencing Bisynthesis (SBS) technology
MiSeq operates on Illumina’s SBS technology, which uses the process of incorporating fluorescently labeled nucleotides into DNA one nucleotide at a time and reading the fluorescent signal to determine the sequence. This process allows DNA sequences to be read with a very high degree of accuracy.
Pair-End Sequence
MiSeq also supports paired-end sequencing, where both ends of a DNA fragment are sequenced for accurate read mapping and detection of structural variation. Paired-end reads can be up to 300 base pairs x 2 reads in length, enabling data acquisition of long reads.
Cluster Formation
In MiSeq, clusters of DNA are formed on a flow cell and the DNA fragments in each cluster are sequenced in parallel. This process provides the basis for next-generation sequencing technologies that can analyze millions of DNA fragments simultaneously.
3. Applications of MiSeq
MiSeq is used in a variety of applications and can be used for a wide range of studies, from small-scale genome analysis to targeted sequencing. The main applications are as follows
3.1 Target sequencing
It is widely used for targeted sequencing to analyze specific genes or genomic regions. It is useful for analysis of cancer gene panels, disease-related genes, and biomarker detection.
3.2 Whole Genome Sequencing
MiSeq also supports whole genome sequencing of small organisms (e.g., bacteria and viruses). Examples include pathogen analysis, evolutionary analysis, and environmental DNA analysis.
3.3 16S rRNA resolution (microbial clump resolution)
MiSeq is widely used for microbiota analysis by 16S rRNA sequencing and is suitable for profiling gut bacteria and microbial communities in the environment. It helps to identify intestinal flora and pathogens.
3.4 Gene expression analysis (RNA sequencing)
MiSeq also supports RNA sequencing, which is used for quantitative analysis of gene expression and detection of splice variants. This allows for the investigation of changes in gene expression associated with disease or therapy.
3.5 Clinical Applications
MiSeq is used for cancer gene panel analysis and genetic disease diagnosis in clinical settings. In particular, mutation analysis by targeted sequencing provides important information in cancer therapy and disease diagnosis.
4. Advantages of MiSeq
- Ease of use: MiSeq is designed to be user-friendly, offering intuitive software and automated workflows. This makes sequencing tasks easy to perform and accessible to researchers with little expertise.
- Rapid analysis: MiSeq can acquire sequence data in a short period of time, making it particularly suited for research and clinical applications that require rapid results.
- High accuracy: Based on Illumina’s SBS technology, the data is extremely accurate. This is very important for variant detection and genetic analysis.
- Cost-effective: MiSeq is ideal for small to mid-size sequencing projects, allowing you to obtain high quality data at a reduced cost.
summary
The MiSeq is Illumina’s next-generation sequencer model for targeted sequencing and small-scale genomic analysis. Featuring a compact design, high flexibility, and rapid data acquisition, the MiSeq is suitable for use in laboratories and clinical settings. the MiSeq is widely used in a variety of fields, including cancer research, microbiological analysis, and genetic disease analysis, as it provides highly accurate data in a short time.
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Supervisor of the article
Dr. Hiroshi Oka
Graduated from Keio University, Faculty of Medicine
Doctor of Medicine
Medical Doctor
