Protocol for ligation of the 3´ OH of RNA to the 5´ pre-adenylated DNA with T4 RNA Ligase 2 truncated (NEB #M0242)

 10X T4 RNA Ligase Reaction Buffer
2 μl
 50% PEG8000*
4 μl
 RNA Substrate (10 μM)
2 μl
 5' pre-adenylated DNA linker (20 μM)
2 μl 
 T4 RNA Ligase 2 truncated
1 μl (200U)
 Nuclease-free water
To 20 μl


Incubate for 1-2 hour at 25°C.

Stop reaction by adding Proteinase K or quenching with EDTA.

* PEG8000 could be added up to 25% in the reaction for boosting ligation efficiency.