Microarray Technology
Microarray technology is a method of greatly increasing the throughput of laboratory processes to efficiently perform large-scale genetic tests, such as comparing gene expression between healthy and tumorous tissues. DNA microarray technology is being used for research on heart disease, mental illness and infectious diseases, as well as for developing targeted cancer treatment strategies.
Performing a microarray analysis requires a machine that can automatically dispense tens of thousands of microscopic dots of DNA fragments in a regular matrix, or array, on the surface of a slide. The slide is called a DNA chip and the deposited DNA spots are called probes, or oligos. The probes are covalently bound to the surface of the slide, or synthesized directly on the slide, and a computer keeps track of which probe is located at each position. Alternatively, the probes are bound to microscopic polystyrene beads.
To prepare the target, DNA samples are purified, treated with reverse transcriptase (RT) enzyme to generate complementary DNA (cDNA), and then coupled with fluorescent or chemiluminescent dye molecules, to allow test results to be read. To perform the microarray analysis, the target cDNA is “hybridized” to the DNA probes on the chip with hydrogen bonds that naturally form between complementary nucleic acid sequences on the target and the probes. The more complementarity, the more strongly the target will bind to each specific probe on the chip.
The relative amounts of cDNA that have been bound to each probe can be “read” by a machine scanning the chip with a laser to illuminate the dye molecules, and then processing the image to detect the presence and strength of dye found at each spot.
Equipment and supplies used for microarray technology includes homogenizers, heating and cooling blocks, and mini-centrifuges to prepare the target; DNA, RNA and protein extraction kits; automated nucleic acid purification equipment; PCR systems to amplify the target; and microscopes and spectroscopes to read the test results.
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