Protocol for ssDNA/RNA Ligation (M0319)

Protocol

  1. Set up the following reaction in a sterile microfuge tube:
  2. Components Volume
    ssDNA/RNA Substrate 20 pmol (1 pmol/ul) 
    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.