TABLE 2.
Analytical gradient mixed species for the separation of mixed 16S rRNA amplicons C (0933F/1407R) at 62.0°C with the WAVE 3500 DNA fragment analysis systema
| Step | Time (mins) | Buffer A (%) | Buffer B (%) | Buffer C (%) |
|---|---|---|---|---|
| Loading | 0.0 | 46 | 54 | 0 |
| Step 1 (5.6 min) | 0.5 | 40 | 60 | 0 |
| Step 2 (5.6 min) | 2.5 | 39 | 61 | 0 |
| Step 3 (5.6 min) | 4.5 | 38 | 62 | 0 |
| Step 4 (5.6 min) | 8.5 | 37 | 63 | 0 |
| Step 5 (5.6 min) | 10.5 | 36 | 64 | 0 |
| Start clean | 10.6 | 46 | 54 | 0 |
| Stop clean | 10.7 | 46 | 54 | 0 |
| Start equilibrate | 10.8 | 46 | 54 | 0 |
| Stop equilibrate | 11.6 | 46 | 54 | 0 |
a
The flow rate was 0.35 mH/min.