| Ligation mix (5 μL per reaction) | Volume (μL) | Final concentration |
|---|---|---|
| Nuclease-free water | 2.25 | |
| T4 ligase (2000 U μL−1) | 1.0 | 80 U μL−1 |
| NEB Cutsmart buffer | 0.5 | 1× |
| ATP (100 mM) | 0.25 | 1 mM (in 25 μL) |
| Methylated adaptor (0.75 μM) | 1.0 | 30 nM (in 25 μL) |
| Total | 5 (total reaction volume: 25 μL) |
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