BsrDI is derived from Bacillus stearothermophilus D70 and consists of two subunits of different sizes, belonging to a heterodimeric enzyme. BsrDI recognizes the specific sequence GCAATG/CATTGC. When cutting the top strand, it cuts at a position 2 nucleotides downstream of the recognition site, while for the bottom strand, it cuts immediately, after the recognition site, generating specific sticky ends. BsrDI belongs to the Type IIS restriction enzymes, which recognize asymmetric DNA sequences and cleave outside of their recognition sequence, making it suitable for Golden gate assembly.

Recommended Reaction Conditions

1× Cut Buffer B；
Incubate at 37℃；
Refer to "Protocol for DNA Digestion" for reaction setup.

Heat Inactivation

Incubation at 80℃ for 20 minutes.

Methylation sensitivity

Cleavage with this restriction enzyme may be blocked or impaired when the substrate DNA is methylated by the EcoBI methylase.

-20℃