Overview of Gene Microarray Chips
Gene microarray chip (Gene microarray chip) is a high-throughput gene analysis platform, which is widely used in the fields of gene expression, gene mutation, gene detection and genomics research.
It is a solid-phase microarray that uses gene sequence fragments as probes to detect thousands to millions of genes or gene expression products simultaneously.
The working principle of gene microarray chips is based on the principle of base recognition by complementary pairing. A large number of DNA fragments or cDNA fragments are immobilized on the surface of the chip, which are usually known gene sequences or probe sequences representing different genes.
The RNA or cDNA in the sample to be examined is labeled and hybridized with the probe sequence on the chip to form a complementary pair. By detecting the signal intensity of the label after hybridization, the relative expression level of each gene in the sample to be examined can be inferred.
The design of gene microarray chips is usually based on two different platforms: whole genome chips and customized chips. Whole genome microarrays contain probes for the entire genome and can be used for comprehensive analysis of all genes.
Customized microarrays, on the other hand, select specific genes or genomic regions to design corresponding probes for specific research purposes according to research needs.
Research goals that can be achieved with gene microarray chips:
- Gene expression analysis: By detecting the expression levels of different genes in the samples, it reveals the regulatory mechanisms of genes under different conditions, gene regulatory networks, and changes in gene expression related to diseases.
- Genetic Variation Testing: Identifies genetic variants associated with disease by detecting gene sequence variants or polymorphisms in samples, and studies the effects of mutations on gene function and disease occurrence.
- Genome analysis: reveals changes in genome structure and function, such as chromosome deletions, copy number variations, etc., by detecting the mutation or methylation status of the entire genome.
- Drug Screening and Individualized Medicine: Screening for potential drug targets or predicting an individual's response to a specific drug by comparing gene expression changes before and after drug treatment.
In conclusion, gene microarray chip is an efficient and high-throughput genetic analysis tool, which is widely used in biomedical research, biotechnology and clinical diagnostics, and provides important data support for revealing gene functions and regulatory mechanisms, studying disease occurrence mechanisms, developing new drugs and realizing individualized medical treatment.
Through the application of gene microarray chips, scientists are able to gain a deeper understanding of the functions and interrelationships of genes in living organisms, which in turn promotes research progress in the fields of genomics and biomedicine.
Advantages of Gene Microarray Chips
- High-throughput analysis: thousands of genes can be detected simultaneously on a single chip, greatly improving research efficiency and data yield.
- High sensitivity: Through the detection of marker signals, the expression level or mutation of genes can be measured with high sensitivity and accuracy.
- Versatility: Gene microarray chips can be applied in many fields such as gene expression analysis, gene mutation detection, genome analysis, etc. and have a wide range of application potential.
- Data integration and analysis: The data output from microarrays can be integrated and analyzed by bioinformatics methods to reveal interactions between genes, signaling pathways and functional patterns.
Gene microarray chips also have some limitations and challenges
- Design dependency: The design of microarrays requires prior determination of the gene probe sequences to be detected, and for unknown genes or newly discovered sequences, additional work is required for design and validation.
- Data processing complexity: The raw data output from the chip is large and complex, requiring data preprocessing, normalization and analysis to extract meaningful biological information.
- Limited coverage: The limited number of probes on the chip may not cover all genes or genomic regions and may not be comprehensive enough for certain research objectives.
- Result verification and validation: the reliability of microarray results needs to be verified by other experimental methods to avoid the effect of false positive or false negative results.
With the continuous development of technology, the popularization of gene sequencing technology and the reduction of cost, some emerging technologies such as RNA sequencing (RNA-seq) have gradually replaced gene microarray chips in some application areas.
However, gene microarray chips still have unique advantages and application space, and remain a valuable tool in specific research scenarios and resource constraints.
Preparation method of gene microarray chip
- Probe design: The gene or genomic region to be studied is first identified and then the corresponding probe sequence is designed. Probes can be DNA fragments, cDNA fragments, or oligonucleotides, and are typically 20 to 70 bases in length.
- Probe synthesis: the designed probe sequence is synthesized. Synthesis can be performed by chemical synthesis methods or photolithography methods based on photomask technology.
- Probe labeling: In order to perform detection on the chip, the probe needs to be labeled. Labeling can be done using different methods such as fluorescent dyes, radioisotopes, biotin, and so on. The choice of labeling depends on the experimental requirements and the detection platform.
- Chip Preparation: The probe is immobilized on the chip surface. Typically, glass or silicon wafers are used as the chip substrate, and the surface is specially treated to improve the immobilization of the probes. The probes can be immobilized on the chip by printing, compositing or jointing.
- Printing method: The probe solution is sprayed, jetted, or embossed onto the chip surface at a predetermined location. The printing method may be inkjet printing, air-jet printing or mechanical printing.
- Synthesis method: probes are chemically synthesized on the chip surface, usually by photolithography or microfluidics. Photolithography methods synthesize probes step by step through light and chemical reactions. Microfluidic technology realizes the synthesis of probes through micro reaction cell and microfluidic control.
- Junction method: the probe is immobilized on the chip by a junction molecule. The junction molecule can be an oligonucleotide, a molecular chain, or a molecular modification group.
- Microarray validation and quality control: The prepared microarray needs to be validated and quality controlled to ensure the quality and reliability of the probes. Common validation methods include probe sequence analysis, hybridization efficiency testing and labeling signal strength detection.
Preparation of gene microarray chips requires specialized equipment and technology, and common chip preparation platforms include Affymetrix, Agilent, Illumina and so on. In addition, the preparation process requires attention to factors such as probe selection, labeling stability, uniformity of the chip surface and chip storage conditions to ensure the accuracy and reproducibility of the chip.
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