Nickase Endonuclease Nt.BbvCI

The optimal reaction temperature is 37℃; CpG-methylated DNA may be inhibited by shearing; EcoBI-methylated DNA may be inhibited by shearing; 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 should be 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.。 Nt. BbvCI is a nicking endonuclease that cleaves only one strand of a double-stranded DNA (dsDNA) substrate ， creating a nick on dsDNA without separating the two strands. Nt. BbvCI is frequently used in isothermal nucleic acid amplification (e. g. ， SDA ， RCA). The DNA nick generated by Nt. BbvCI triggers the strand displacement reaction of polymerase ， and repeated cycles of cleavage ， displacement and extension result in exponential amplification of nucleic acids. Concentration: 5?U/μLRecognition site: CCTCAGC5 ‘…CC↓TCAGC…3 ‘3 ‘…GGAGTCG…5 ‘Definition of enzyme activity:One unit (U) is defined as the amount of enzyme required to completely convert 1?μg of supercoiled p615 DNA into the open-circular form in a 50?μL reaction system at 37?°C within 1 hour.