Dpn I digestion

Purpose

To digest (Adeno) methylated GATC sites. Useful for removing cell-derived plasmid template from PCR samples.

Procurement

Can be ordered directly from NEB.
Typically stored at -20°C; can be found in the common enzyme freezer box.

Use

Use in NEBuffer 4; however, it can be added directly to PCR sample.

Incubate at 37°C for one hour or overnight.
Heat inactivate by incubating at 80°C for 20 minutes.

NEB 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.

NEB buffer composition:

1X NEBuffer 4:
20 mM Tris-acetate
50 mM potassium acetate
10 mM Magnesium Acetate
1 mM Dithiothreitol
pH 7.9 @ 25°C