Klenow Reaction

Overview

This protocol describes how to blunt ends of DNA by 3'Protocol for blunting ends by 3' overhang removal and 3' recessed end fill-in.

Protocol

• DNA should be dissolved in 1X NEBuffer 1-4 or T4 DNA Ligase Reaction Buffer
• Supplement with 33 μM each dNTP.
• Add 1 unit Klenow per microgram DNA and incubate 15 minutes at 25°C.
• Stop reaction by adding EDTA to a final concentration of 10mM and heating at 75°C for 20 minutes.