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.
Application
TaqI is a restriction endonuclease used in molecular biology applications to cleave DNA moledcules at the recognition site 5′-T/CGA-3′, generating fragments with 5′-cohesive ends.
Other Notes
Supplied with 10x Restriction Enzyme 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.
Physical form
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
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