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Tagging pathogens with synthetic DNA 'barcodes'

NA that carries the genetic code in living cells consists of two complementary strands that attach to one another along their length. But Luo's research purposely and completely ignores the DNA's genetic coding properties. He uses DNA, he said, as a "generic instead of a genetic material."

By synthesizing three short strands of DNA, each of which is complementary to one of the others along half its length, the researchers can create a Y-shaped structure. Combining several of these structures creates a web with many branching ends. "While DNA is flexible, the short strands used here are quite rigid," Luo said. "A long piece of spaghetti is floppy, but a short bit of it is quite stiff."

An antibody or some other molecule that will bind to the molecule to be detected is attached to one of the loose ends of the DNA. To other ends are attached molecules of fluorescent dye in a predetermined pattern.

For example, one probe might contain four molecules of green dye and one of red. Another might have three molecules of green and two of red, and so on. If a mixture of several probes is added to a solution containing, for example, E. coli bacterial DNA, only probes with a particular color code will be programmed to bind to that DNA. The results can be seen under a fluorescent light microscope using colored filters that pass only one color at a time. A signal in which the ratio of intensity of green light is four times that of red light, for example, identifies a "4G1R" probe. The researchers say that up to 1,000 different codes can be created using only three fluorescent dyes.

To amplify the signals, the researchers attached many DNA probes to the surface of polystyrene microbeads 5.5 microns (millionths of a meter) in diameter. The results can be read in several ways. One is in a flow cytometer, in which samples move rapidly past a window where a computer reads the color codes of individual beads. Another is by dot blotting, in which the sampl
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Source:Cornell University News Service


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