T5 Exonuclease degrades DNA in the 5' to 3' direction. T5 Exonuclease is able to initiate nucleotide removal from the 5' termini or at gaps and nicks of linear or circular dsDNA. However, the enzyme does not degrade supercoiled dsDNA. T5 Exonuclease also has ssDNA endonuclease activity.

• Double-stranded DNA specific exonuclease and single-stranded DNA endonuclease
• Initiates at the 5' termini of linear or nicked double-stranded DNA
• Cleaves linear or nicked double-stranded DNA in the 5' to 3' direction