Protocol for Oligonucleotide Adenylation (NEB #E2610)


  1. Set up the following reaction in a sterile microfuge tube:
    Components Volume
    Phosphorylated DNA
    100 pmol (5 pmol/µl)
    10X 5´ DNA Adenylation
    Reaction Buffer
    2 µl
    1 mM ATP 2 µl
    Mth RNA Ligase 2 µl (100 pmol)
    Nuclease-free Water to 20 µl
  2. Incubate at 65°C for 1 hour.
  3. Inactivate the enzyme by incubation at 85°C for 5 minutes.