Tris HCl, pH 7.5 supplemented with 1 mM αKG, 1 mM L-Asc, and 75 μM Fe(II) is the optimal reaction buffer for FTO RNA Demethylase (86% conversion of m6A).
The relative activity of FTO RNA Demethylase with NEB Buffers supplemented with 1 mM αKG, 1 mM L-Asc, and 75 μM Fe(II), results in the following % conversion of m6A.
| Buffer | % Conversion |
|---|---|
| 50 mM Tris HCl, pH 7.5 | 86 |
| rCutSmartTM Buffer | 24 |
| NEBuffer 1.1 | 73 |
| NEBuffer 2.1 | 24 |
| NEBuffer 3.1 | 4 |
