Restriction Endonuclease BspEI

The enzyme volume added to the reaction system should not exceed 10% of the total volume to avoid excessive glycerol in the enzyme causing asterisk activity;
The additives (e.g., glycerol, salt) in the restriction endonuclease storage buffer are the same as the 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;
Cleavage may be inhibited for DNA methylated with Dam;
Cleavage may be inhibited for DNA methylated with CpG;
No asterisk activity was observed after 3 h incubation, but asterisk activity may appear after delayed digestion;
Isoschizases may have different sensitivities to different methylation modifications;
Inactivation conditions: Incubation at 80℃ for 20 min. BspEI is a Type IIP restriction enzyme derived from Bacillus species , obtained through recombinant expression and purification in Escherichia coli. BspEI recognizes and cleaves the sequence TCCGGA , generating a 4-base 5 ' sticky end. BspEI requires a dedicated buffer and is incompatible with the Reaction Buffer of the QuickCut series. Concentration: 10?U/μLIsoschizomers: Kpn2I , MroI , AccIII , Aor13HI , Bsp13I , BseAIRecognition site: TCCGGA5 '…T↓CCGGA…3 '3 '…AGGCC↑T…5 ' Definition of enzyme activity:One unit (U) is defined as the amount of enzyme required to completely digest 1?μg of λDNA (Dam?) in a 50?μL reaction system at 37?°C within 1 hour.