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  • Protocol for ssDNA/RNA Ligation (M0319)

    Protocol

    1. Set up the following reaction in a sterile microfuge tube:
    2. Components Volume
      ssDNA/RNA Substrate 6–8
      5´ App DNA Oligonucleotide 40 pmol (2 pmol/µl)
      10X NEBuffer 1 2 µl
      50 mM MnCl2 (for ssDNA ligation only) 2 µl
      Thermostable 5´ App DNA/RNA Ligase 2 µl (40 pmol)
      Nuclease-free Water to 20 µl
    3. Incubate at 65°C for 1 hour.
    4. Inactivate the enzyme by incubation at 90°C for 3 minutes.