Restriction Endonuclease BsrDI

The enzyme volume added to the reaction system should not exceed 10% of the total volume to avoid excessive glycerol in the enzyme causing star-shaped activity; Additives (e.g., glycerol, salt) in the restriction endonuclease storage buffer are the same as contaminants (e.g., salt, EDTA, or ethanol) in the substrate solution; the smaller the reaction volume, the stronger the inhibitory effect of the enzyme digestion reaction; BsrDI can be used in QuickCut series reaction beffers, but star-shaped activity is significantly enhanced. If using QuickCut series reaction beffers, it is not recommended to digest for more than 3 hours or more than 2 U of BsrDI/ug of DNA substrate; Shearing may be inhibited for DNA methylated by EcoBI; Isolytic enzymes may have different sensitivities to different methylation modifications; Inactivation conditions: incubate at 80℃ for 20 min. BsrDI is derived from Bacillus stearothermophilus D70. It consists of two subunits of different sizes and is a heterodimeric enzyme. BsrDI recognizes the specific sequences GCAATG/CATTGC. It cleaves the top strand 2 nucleotides downstream of the recognition site ， while the bottom strand is cleaved immediately after the recognition site ， generating specific sticky ends. As a Type IIS restriction enzyme ， it can be used for Golden Gate assembly. Concentration: 10?U/μLIsoschizomers: Bse3DI ， BseMIRecognition site: GCAATGNN5 ‘…GCAATGNN↓…3 ‘3 ‘…CGTTAC↑NN…5 ‘Definition of enzyme activity:One unit (U) is defined as the amount of enzyme required to completely digest 1?μg of λDNA in a 50?μL reaction system at 37?°C within 1 hour.