NEBNext dA-Tailing Module Protocol (E6040)

Introduction

Starting Material: 1–5 μg of end repaired, blunt DNA (100–1000 bp)in 37 μl H₂O.

Protocol

1. Mix the following components in a sterile microfuge tube:
   
   End Repaired, Blunt DNA:   42 μl
   NEBNext dA-Tailing Reaction Buffer (10X):   5 μl
   Klenow Fragment (3´→ 5´ exo^–):   3 μl
   -------------------------------------------------------------
   Total volume:   50 μl
2. Incubate in a thermal cycler for 30 minutes at 37°C.
3. Purify DNA sample on one column and elute in 25 μl of sterile dH₂O or elution buffer.