Unveiling the Genome: What is Microarray Analysis? - Healthcare-netizens/arpita-kamat GitHub Wiki
Imagine a tiny chip, no bigger than a postage stamp, capable of simultaneously analyzing the activity of thousands, even millions, of genes. This revolutionary technology is microarray analysis, a powerful tool that has transformed our understanding of genomics, disease, and biological processes. At its core, microarray analysis is a high-throughput method that allows researchers to measure gene expression levels, detect genetic variations, or identify specific DNA sequences on a massive scale.
The fundamental principle behind microarray technology is nucleic acid hybridization. This biological phenomenon describes the ability of two complementary single-stranded nucleic acid molecules (DNA or RNA) to bind together, forming a stable double helix. A microarray chip, typically a glass slide or silicon wafer, is essentially a precisely organized collection of microscopic spots, each containing a unique DNA sequence called a "probe." These probes are designed to be complementary to specific target genes or genomic regions of interest.
The process typically begins with the isolation of nucleic acids (DNA or RNA) from a biological sample – this could be cells from a healthy individual, a diseased tissue, or even a cell line treated with a new drug. For gene expression studies, messenger RNA (mRNA) is usually isolated, as it represents the genes actively being transcribed and translated into proteins. This mRNA is then reverse-transcribed into complementary DNA (cDNA) and labeled with a fluorescent dye. In many experiments, two samples (e.g., a "test" sample and a "control" sample) are labeled with different fluorescent dyes (e.g., Cy3 for green and Cy5 for red) to allow for direct comparison on the same chip.
The labeled cDNA is then hybridized to the microarray chip. During hybridization, the labeled DNA fragments from the sample will bind, or "hybridize," only to their complementary probes on the chip. After a washing step to remove unbound material, the chip is scanned by a specialized laser scanner. The scanner excites the fluorescent dyes, and the emitted light is detected, creating an image of the chip. The intensity of the fluorescence at each spot is directly proportional to the amount of labeled DNA that hybridized to that specific probe, thereby indicating the abundance of the corresponding gene or sequence in the original sample.
The resulting data is a vast collection of intensity values, which are then analyzed using sophisticated bioinformatics software. This analysis allows researchers to identify which genes are "turned on" (upregulated), "turned off" (downregulated), or show no change in expression between different conditions. Beyond gene expression, microarrays can also be designed to detect single nucleotide polymorphisms (SNPs), copy number variations (CNVs), or even identify specific pathogens, making them incredibly versatile tools in genetics, medicine, and beyond.
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