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The two strands of DNA have opposite chemical polarity which means that their sugar-phosphate backbones run in opposite directions. The direction is specified by referring to the carbons of the ribose ring in the sugar-phosphate backbone of DNA. 5' specifies the 5th carbon in the ribose ring, counting clockwise from the oxygen molecule, and 3' specifies the 3rd carbon in the ring. Direction of, and in reference to, DNA molecules is then specified relative to these carbons. For example, transcription, the act of transcribing DNA to RNA for eventual expression, always occurs in the 5' to 3' direction. Nucleic acid polymerization cannot occur in the opposite direction, 3' to 5', because of the difference in chemical properties between the 5' methyl group and the 3' ring-carbon with an attached hydroxyl group.

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