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  • Protocol for Oligonucleotide Adenylation (E2610)

    Protocol

    1. Set up the following reaction in a sterile microfuge tube:
      Components Volume
      Phosphorylated DNA
      Oligonucleotide
      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.