Description: T5 exonuclease was purified from E. coli strain BL21(DE3) carrying a plasmid overexpressing His-tagged T5 exonuclease by using a Ni-NTA column and a Q-Sepharose FF column. The N-terminal His-tag does not affect its activities. A 10% SDS PAGE gel shows the purity of T5 exonuclease. T5 exonuclease is stored in the storage buffer. T5 exonuclease is free from RNase and other DNase contaminations.
T5 exonuclease is a highly processive 5′→3′ exonuclease that degrades linear and circular single-stranded DNA (ssDNA), and linear, nicked, and gapped double-stranded DNA (dsDNA) from its 5′ ends or at nicks and gaps. It can digest any linear DNA including dogged-bone DNA. T5 exonuclease has flap endonuclease (FEN) activities. As a result, it can completely digest supercoiled circular DNA forming a hairpin structure, such as supercoiled plasmid pAB1. T5 exonuclease does not digest relaxed and supercoiled circular DNA, such as pUC18 and pBR322 that does not form hairpin structures.
Concentrations: Two concentrations of T5 exonuclease are available: 1 μM (37.5 μg/mL) and 5 μM (187.5 μg/mL). Typically, 10 to 50 nM of T5 exonuclease at 37 °C for 30 min is recommended and sufficient for most assays. For completely digesting DNA into nucleotides, 200 nM of T5 exonuclease at 37 °C for 2 hours should be used.
Shipping and storage: T5 exonuclease is shipped with ice gel packs and should be stored in a -20°C freezer upon arrival and is stable for 12 months in this condition from the date of receipt.
Dilution buffer: 50 mM Tris-HCl, pH 8.0, 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, and 50% glycerol. Dilution buffer is supplied.
10xT5 exonuclease assay buffer: 500 mM Tris-HCl, pH 8.0, 660 mM KAC, 100 mM Mg(AC)2, and 5 mM DTT. 10xassay buffer is supplied.
T5 exonuclease is a highly processive 5′→3′ exonuclease that degrades linear and circular single-stranded DNA (ssDNA), and linear, nicked, and gapped double-stranded DNA (dsDNA) from its 5′ ends or at nicks and gaps. It can digest any linear DNA including dogged-bone DNA. T5 exonuclease has flap endonuclease (FEN) activities. As a result, it can completely digest supercoiled circular DNA forming a hairpin structure, such as supercoiled plasmid pAB1. T5 exonuclease does not digest relaxed and supercoiled circular DNA, such as pUC18 and pBR322 that does not form hairpin structures.
Concentrations: Two concentrations of T5 exonuclease are available: 1 μM (37.5 μg/mL) and 5 μM (187.5 μg/mL). Typically, 10 to 50 nM of T5 exonuclease at 37 °C for 30 min is recommended and sufficient for most assays. For completely digesting DNA into nucleotides, 200 nM of T5 exonuclease at 37 °C for 2 hours should be used.
Shipping and storage: T5 exonuclease is shipped with ice gel packs and should be stored in a -20°C freezer upon arrival and is stable for 12 months in this condition from the date of receipt.
Dilution buffer: 50 mM Tris-HCl, pH 8.0, 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, and 50% glycerol. Dilution buffer is supplied.
10xT5 exonuclease assay buffer: 500 mM Tris-HCl, pH 8.0, 660 mM KAC, 100 mM Mg(AC)2, and 5 mM DTT. 10xassay buffer is supplied.
This product is used for research only
T5 exonuclease assay instruction: