BsaI

Recognition site:
GGTCTC(N)₁Λ
CCAGAG(N)₅

Source:
An E. coli strain that carries the BsaI gene from  Bacillus stearothermophilus

Unit Definition
One unit is defined as the amount of enzyme required to digest 1 µg of pXba DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Condition
1X BsaI Buffer at 37°C

10x BsaI Buffer:
100 mM Tris-HCl (pH 8.5 at 37°C), 100 mM MgCl_2, 1 M KCl, 1 mg/ml BSA

Storage Buffer:
10 mM Tris-HCl, 300 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 500 μg/ml BSA, 50% Glycerol, pH 7.4 @ 25°C

Heat Inactivation
65°C for 20 min

Typical ligation/recut assay result:
More than 95% of the DNA fragments can be ligated after a 10-fold over-digestion of pXba DNA with BsaI, and more than 95% of these DNA fragments can be recut with BsaI.

Recommended storage condition: -20°C

Comments:
Blocked by overlapping dcm methylation. Cleavage of mammalian genomic DNA is blocked by some combinations of overlapping CpG methylation. Activity at 50°C is 100%.