DpnI

Description

Recognition site: G(CH3)A^TC

Source:
An E. coli strain that carries the DpnI gene from Diplococcus pneumonia

Supplied With:
10x DpnI Buffer

10x DpnI Buffer:
500 mM Potassium Acetate
200 mM Tris-acetate
100 mM Magnesium Acetate
1 mg/ml BSA
pH 7.9 @ 25°C

Supplied In:
10 mM Tris-HCl, 300 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 500 μg/ml BSA, 50% Glycerol

Recommended Storage Condition: -20°C

Typical ligation/recut assay results:
More than 70% of the pBR322 DNA fragments can be ligated and more than 95% of these can be re-cut after 50-fold over-digestion with DpnI.

Unit Definition：
One unit is defined as the amount of enzyme required to digest 1 µg of pBR322 DNA (dam methylated) in 1 hour at 37°C in a total reaction volume of 50 µl.

Comments:
Resistant to heat inactivation (20 minutes, 85°C). DpnI requires the presence of N6-methyladenine within the recognition sequence to cleave DNA. DNA purified from a dam+ strain will be a substrate for DpnI. DpnI will only cleave fully-adenomethylated dam sites. Hemi-adenomethylated dam sites DpnI cleaves 60X more slowly.

Additional information

Manual & Protocols

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