|
| |||
|
Amount (μl)
|
|||
| Component | C and 5-mc test | 5-hmc test | Final concentration |
|
| |||
| PfuTurbo Cx reaction buffer (10×) | 5 | 5 | 1× |
| dNTP mix (25 mM each) | 0.4 | 0.4 | 200 μM each |
| 5-mC_test_F (10 μM) | 1 | — | 0.2 μM |
| 5-mC_test_R (10 μM) | 1 | — | 0.2 μM |
| 5-hmC_test_F (10 μM) | — | 1 | 0.2 μM |
| 5-hmC_test_R (10 μM) | — | 1 | 0.2 μM |
| Bisulfite-treated DNA (from Step 19) | 2 | 2 | |
| PfuTurbo Cx DNA Polymerase (2.5 U μl−1) | 1 | 1 | 2.5 U per 50 μl PCR |
| Milli-Q water | to 50 | to 50 | |
| Final volume | 50 | 50 | |
|
| |||
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