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) |