Table 1.
Listed are the steps taken in the polymerase chain reaction (PCR) protocol followed by the primers used during PCR (Forward and Reverse) and subsequent pyrosequencing (Sequencing) in order to generate targeted analysis of OXTR CpG site -924.
| Step | Process | Temperature (°C) | Duration (s) | Cycle(s) |
|---|---|---|---|---|
| 1. | Initial denaturation | 95 | 15 | 1 |
| 2. | Denaturation | 94 | 30 | 50 |
| Annealing | 56 | 30 | ||
| Elongation | 72 | 30 | ||
| 3. | Elongation | 72 | 10 | 1 |
| 4. | Hold | 4 | until analysis | 1 |
| Primer | Sequence |
|---|---|
| Forward (TSL101F) | 5′-TTGAGTTTTGGATTTAGATAATTAAGGATT-3′ |
| Reverse (TSL101R) | 5′-biotin-AATAAAATACCTCCCACTCCTTATTCCTAA-3′ |
| Sequencing (TSL101S) | 5′-AGAAGTTATTTTATAATTTTT-3′ |