NEBNext dA-Tailing Module Protocol (E6053)


Introduction

Starting Material: 1-5 μg of of end repaired, blunt DNA (100-1000 bp).

Protocol

  1. Mix the following components in a sterile microfuge tube:
     End Repaired, Blunt DNA variable 
     NEBNext dA-Tailing Reaction Buffer (10X)  5 μl
     Klenow Fragment (3´→ 5´ exo)  3 μl
     Sterile H2O  variable
     Total volume  50 μl
  2. Incubate in a thermal cycler for 30 minutes at 37°C with the heated lid set to ≥ 45°C.
  3. Purify DNA sample on one spin column or using AMPure XP beads.

Note: for details how this module is used in the NEBNext Library Prep for Illumina workflow, please see manual for NEBNext DNA library Prep Master Mix Set for Illumina (NEB #E6040).