Restriction Endonuclease ApeKI

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; The optimal reaction temperature is 75℃; For CpG-methylated DNA, cleavage may be hindered and it cannot be heat-inactivated; No asterisk activity was observed after 3 hours of incubation, but asterisk activity may appear after delayed digestion; Isolytic enzymes may have different sensitivities to different methylation modifications. ApeKI is derived from the hyperthermophilic aerobic archaeon Aeropyrum pernix K1 and belongs to the Type IIP restriction enzymes. It recognizes and cleaves the sequence G/CWGC, generating sticky ends with a 3-base overhang. ApeKI is one of the preferred enzymes for genotyping-by-sequencing (GBS). In addition, it is widely used in fields such as genetic research of animals and plants, population evolutionary analysis , and functional gene mapping. Concentration: 10 U/μLisoschizomers: TseIRecognition site: GCWGC5 '…G↓CWGC…3 '3 '…CGWC↑G…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 75 °C within 1 hour.