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Cat. No.: DpnI-500 (for 500U)

Cat. No.: DpnI-2500 (for 2500U)

Cat. No.: DpnI-10k (for 10KU)

Cat. No.: DpnI-50k (for 50KU)

Description

DpnI recognizes and cleaves the GATC sequence in DNA double strands where adenine at the N6 position is methylated (N6-methyladenine). When both adenines at the N6 position within the recognition site of DpnI are methylated, the enzyme exhibits normal activity. However, if only one adenine at the N6 position is methylated, the enzyme activity is reduced by approximately 60-fold.

Plasmids extracted from dam+ (positive for Dam methylase expression) Escherichia coli strains such as DH5α contain GATC sequences with adenine N6 methylation, making them susceptible to recognition and cleavage by DpnI. On the other hand, plasmids extracted from dam- (negative for Dam methylase expression) E. coli strains such as JM110 lack adenine N6 methylation within the GATC sequence, rendering them unrecognizable and uncuttable by DpnI.

Enzyme storage buffer 

10 mM Tris-HCl (pH 7.4 at 25°C), 300 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.5 mg/ml BSA, and 50% glycerol.

Enzyme digestion and ligation efficiency

Digestion with a 50-fold excess of the endonuclease for 1 hour results in >70% of the digested pBR322 DNA fragments being able to be ligated. Furthermore, >95% of these fragments can be recut by the enzyme.

Unit activity

One unit (1 U) of the enzyme activity is defined as the amount of enzyme required to completely digest 1 μg of λDNA in a 50 μl reaction system at 37°C for 1 hour.

Precautions

• The endonuclease should be stored in an icebox or on an ice bath during use and promptly placed at -20°C after each use for storage.
• If the expected cleavage sites are not cut, please verify if the specific sites have been methylated.
• This product is intended for scientific research purposes by professionals only and should not be used for clinical diagnosis or treatment. It should not be used for food or drug purposes and should not be stored in residential areas.
• For your safety and health, please wear laboratory attire and disposable gloves when handling the product.

Storage

The minimum shelf life is 2 years at -20°C.