Table 2.
Background cleavage by GlaI in a coupled M.SssI assay
| Concentration of oligonucleotide 1 (µM) | Rate of fluorescence change (AU/s) |
Background cleavage (%) | |
|---|---|---|---|
| Assay | Negative control | ||
| 0.03 | 3.33 | 0.47 | 14.1 |
| 0.05 | 4.07 | 0.38 | 9.34 |
| 0.08 | 5.36 | 0.59 | 11.0 |
| 0.13 | 7.53 | 0.88 | 11.7 |
| 0.22 | 10.4 | 1.25 | 12.0 |
| 0.36 | 14.3 | 1.48 | 10.4 |
| 0.60 | 18.4 | 1.64 | 8.91 |
| 1.00 | 21.8 | 0.30 | 1.38 |
Assays contained oligonucleotide 1, AdoMet, M.SssI and GlaI. Negative control assays were prepared identically except that they lacked M.SssI. Background cleavage was calculated as the percentage of assay signal caused by non-specific cleavage of partially methylated oligonucleotide 1 by GlaI.