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

NEB unit definition:

One unit is defined as the amount of enzyme required to digest 1 g of pBR322 DNA (dam methylated) in 1 hour at 37C in a total reaction volume of 50 l.

NEB buffer composition:

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


This topic: Lab > WebHome > ProtocolList > ProtocolUpdatesNeeded > ProtocolsDpnIDigestion
Topic revision: r1 - 2012-08-15 - CraigBarnhart