End Repair of DNA Protocol - NEBNext Fast DNA Library Prep Set for Ion Torrent (E6270)

Overview

Starting Material: 10 ng–1 μg of Fragmented DNA.

Note: For use with the Ion Xpress™ Barcode Adapters 1-16 Kit, a minimum of 100 ng starting material is recommended. Lower amounts may cause adaptor concatamerization.

Protocol

  1. Mix the following components in a sterile microfuge tube on ice:
    Fragmented DNA   1–51 μl
    NEBNext End Repair Reaction Buffer   6 μl
    NEBNext End Repair Enzyme Mix   3 μl
    Sterile H2O   variable
    -------------------------------------------------------------------------
    Total volume   60 μl

  2. Incubate in a thermal cycler for 20 minutes at 25°C, followed by 10 minutes at 70°C.

  3. Pulse spin the microfuge tube and return to ice.