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Dpn I
Neoschizomers: BfuC I, Bsp143 I, BstEN II, BstMB I, BstKT I, Dpn II, Kzo9 I, Nde II | GA↓TC | |
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S pack |
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EN-160S |
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200 Units |
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25,00 |
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L pack |
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EN-160L |
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1000 Units |
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100,00 |
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| | Source: An E. coli strain that carries the dpnIR gene from Diplococcus pneumoniae G41.
Buffer supplied: 10x Dpn I (BSA included).
Substrate for unit definition: pBR322 (22 sites)
Reaction conditions: 66 mM potassium acetate, 33 mM Tris-acetate (pH 7.9 @ 25°C), 10 mM magnesium acetate, 100 µg/ml BSA.
Incubate at 37°C.
| | Storage buffer: 400 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml BSA, and 50% glycerol. Store at -20°C.
Ligation and recutting: After 50-fold overdigestion with Dpn I, >70% of the DNA fragments can be ligated and >95% of these can be recut.
Note: Dpn I is specific for methylated and hemimethylated DNA. Since DNA isolated from most E. coli strains is dam methylated, it is susceptible to Dpn I digestion. Hence, Dpn I is frequently used after a PCR reaction to digest the methylated parental DNA template and select for the newly synthesized DNA containing mutations.
Heat inactivation: 80°C for 20 minutes. | | |
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