Guidelines for ligation of single-stranded pre-adenlyated DNA adapters to the 3’-ends of single-stranded RNA

Protocol

  1. Mix the following reagents:
    10 pmoles ssRNA
    20 pmoles pre-adenylated DNA adapter (e.g. NEB #S1315)
    1x T4 RNA Ligase 2 truncated K227Q reaction buffer
    200 U T4 RNA Ligase 2 truncated K227Q
    20 μl total volume
  2. Incubate reactions at 16°C overnight.