Table 3.
Suggested pretreatment and extraction protocol.
| Step | Time |
|---|---|
| Thaw sample on ice | >1 h |
| 300 mg 0.1 mm silica beads/tube (BioSpec Products Cat# 110 79101) | |
| 100–150 mg sample/tube | |
| 1 mL ASL stool lysis buffer/tube (Qiagen Cat# 19082) | |
| Vortex | 1 min |
| Heat shock 95°C | 5 min |
| Heat shock incubation on ice | 5 min |
| Bead beating 5000 rpm | 3 × 20 s |
| Incubation on ice between each bead beating set | 1 min |
| Pellet beads by centrifugation at 2500 g | 1 min |
| Load 200 μL supernatant/sample into the QIAamp cador Pathogen Kit | |
| Follow the QIAamp cador Pathogen Kit Protocol with the following adjustments: | |
| ● Proteinase K treatment at 70°C | 10 min |
| ● Elute the extract in 100 μL of the included AVE buffer/tube |