Restriction Endonuclease BbvCI

For CpG-methylated DNA, cleavage may be inhibited; 3-hour incubation did not show asterisk activity, but longer digestion may result in asterisk activity; 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; 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; Inactivation conditions: incubation at 80℃ for 20 min. BbvCI is obtained through recombinant expression in Escherichia coli from the BbvCI gene of Bacillus brevis (L. Ge). BbvCI is a Type IIA restriction enzyme that exists as a heterodimer composed of two distinct catalytic subunits, each with an independent cleavage site. BbvCI recognizes and cleaves the non-palindromic sequence CCTCAGC (?5/?2), generating sticky ends with a 3-base overhang at the 5 ' end. BbvCI uses the Reaction Buffer of the QuickCut series and can be used for double digestion with other QuickCut restriction enzymes. Prolonged digestion (over 3 hours) is not recommended to avoid star activity. Concentration: 2 U/μLRecognition site: CCTCAGC5 '…CC↓TCAGC…3 '3 '…GGAGT↑CG…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.