T5 Exonuclease is a nucleic acid exonuclease that digests DNA from the 5' end in the 5'→3' direction for both double-stranded and single-stranded DNA. It is capable of digesting DNA at nicks or gaps in linear or circular double-stranded DNA,however, it cannot  digest supercoiled DNA. In addition, when the Mg²⁺ concentration in the solution is below 1 mM, the activity of T5 Exonuclease towards single-stranded DNA is inhibited. Consequently, T5 Exonuclease is particularly suitable for degrading linearized and nicked plasmids, removing non-circular DNA from ligation products, seamless cloning (Gibson Assembly), and other applications.
 

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