3´ End Labeling of RNA using T4 RNA Ligase 1


Set up the labeling reaction at room temperature until the ligase is added to avoid DMSO precipitation. The addition of DMSO is essential.


  1. Set up a 30 μl reaction as follows:
    1 X Reaction Buffer (50 mM Tris-HCl, pH 7.8, 10 mM MgCl2, 1 mM DTT)
    1mM ATP
    1 μg RNA
    10 % (v/v) DMSO
    1 μM [32P]pCp
    1 μl (10 units) T4 RNA Ligase
  2. Incubate overnight at 16°C