Taq I from Thermus aquaticus 

Supplied with 10x Restriction Endonuclease Buffer SB (B8781).

Comment: Taq I will only partially cleave DNA isolated from E. coli strains that have the dam methylase (dam⁺ strains). Taq I is inefficient in digesting single stranded DNA . Overlapping dam methylation will block cleavage by Taq I. Activity of the enzyme is enhanced by BSA and higher temperatures, >37 °C. Optimal temperature is 65 °C.

Recognition sequence: 5′-T/CGA-3′
Ligation and recutting results: After 2-10-fold Taq I overdigestion of 1 μg λ DNA substrate, results in >95% cutting, >85% of fragments can be ligated, and >95% recut.
Heat inactivation: 80 °C for 20 minutes.

Solution in 20 mM Tris-HCl, pH 8.0, 1 mM EDTA, 100 mM NaCl, 300 mM KCl, 7 mM 2-mercaptoethanol, 50% glycerol (v/v) at 4 °C

expressed in Escherichia coli (Strain that carries a Taq I overproducing plasmid.)

buffered aqueous glycerol solution

10,000 units/mL

wet ice

−20°C

Kessler, C., et al., Specificity of restriction endonucleases and DNA modification methyltransferases: a review (edition 3). Gene 92, 1-248, (1990) abstract

Gruenbaum, Y., Restriction enzyme digestion of hemimethylated DNA. Nucleic Acids Res. Nucleic Acids Res. 9, 2509-2515, (1981) abstract

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LIG2, QuickLink^™ DNA Ligation Kit

D2886, DNA Ligase from T4-infected Escherichia coli

B8781, Restriction Endonuclease Buffer SB