Guidelines for ligation of single-stranded pre-adenlyated DNA adapters to the 3’-ends of single-stranded RNA
- 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
- Incubate reactions at 16°C overnight.