T5 Exonuclease

T5 Exonuclease is a highly efficient 5´→3´ exonuclease for either ssDNA or dsDNA. It also has ssDNA specific endonuclease activity in the presence of magnesium ions. However, the enzyme does not degrade supercoiled dsDNA. The mode of action of T5 Exonuclease in vivo may be analogous to that of the 5´→3´ exonuclease activity of E. coli DNA Polymerase I.

Product Source:

• E. coli strain expressing the T5 phage D15 gene.

Product Includes:

• T5 Exonuclease
• T5 Exonuclease Reaction Buffer [10X]

Applications:

• Plasmid mutagenesis methods.
• Oligonucleotide site-directed mutagenesis.
• Generation of plasmid-sequencing templates.
• Generation of 3’-overhang for improved cloning procedures.

 T5 Exonuclease reaction buffer [10X] composition:

330 mM Tris-acetate (pH 7.5), 660 mM potassium acetate, 100 mM magnesium acetate, 5.0 mM DTT.

Storage Buffer:

50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25ºC

Storage Temperature:

-20ºC

Unit Definition:

One unit of T5 Exonuclease catalyzes the release of 1 nmol of acid-soluble nucleotides from double-stranded calf thymus DNA in 30 minutes at 37°C under standard assay conditions.