| Reagent | Volume (μl) per reaction | Final concentration |
|---|---|---|
| Nuclease-free distilled water | 252.5 | |
| 10× RNA Ligase Buffer (supplied with T4 RNA ligase 2, truncated KQ) | 50 | 1× |
| Annealed linker (from Step 10) | 25 | ~0.4 µM |
| RiboLock RNase inhibitor | 12.5 | 1 U/µl |
| Total | 330 |
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