| Reagents | Volume (μL) | Final |
| PNK reaction | 100 | |
| 10x T4 RNA ligase buffer | 6 | 1x |
| 10 mM ATP | 6 | 1 mM |
| Rnase inhibitor (20 U/µL) | 4 | 0.5 U/µL |
| T4 Rnl1 (10 U/µL) | 40 | 2.5 U/µL |
| Nuclease free water | 4 | |
| Total | 160 |
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