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3' to 5' orientation the direction along a Polynucleotide molecule as defined by proceeding from the 3´-hydroxyl terminus towards the 5´-phosphate terminus. This is the direction opposite to that in which DNA and RNA are synthesized enzymatically. An example of an enzyme which proceeds with this orientation is a 3'-5' exonuclease (a "proofreading" exonuclease), which can assist in correcting errors introduced during DNA replication.

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