Restriction Endonuclease SgrAI

For CpG-methylated DNA, cleavage may be inhibited; for EcoKI-methylated DNA, cleavage may be inhibited; for EcoBI-methylated DNA, cleavage may be inhibited; digestion requires two or more recognition sites; the amount of enzyme used per μg of DNA substrate should not exceed 10 U; due to enzyme stability during the reaction, even incubation time exceeding 1 h will not improve digestion efficiency unless the amount of enzyme is increased; no asterisk activity was observed after 3 h of incubation, but asterisk activity may appear after longer digestion times; prolonged digestion time, high enzyme concentrations, or glycerol concentrations >5% may produce asterisk activity, and overnight digestion is especially not recommended; inactivation conditions: incubation at 80°C for 20 min. SgrAI is derived from Streptomyces griseus and is purified after recombinant expression in Escherichia coli. It specifically recognizes and cleaves the sequence CRCCGGYG. SgrAI is a restriction enzyme not included in the QuickCut series, but it is compatible with Reaction Buffer and can be used for double digestion together with other QuickCut restriction enzymes. Due to its unique enzymatic properties, SgrAI requires two or more recognition sites for efficient cleavage; its cleavage efficiency is significantly reduced for substrates containing only a single site. Concentration: 10 U/μLRecognition site: CRCCGGYG5 '…CR ↓ CCGGYG…3 '3 '…GYGGCC ↑ RC…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.