Common issues & fixes
| Symptom | Likely cause | Fix |
|---|---|---|
| Low/variable recovery | Carrier RNA omitted; incomplete lysis | Add carrier RNA for low-titer inputs; extend lysis mixing. |
| Inhibition in RT-qPCR | Wash carryover | Add/extend washes; ensure full supernatant removal. |
| Cross-contamination | Aerosols / tip reuse | Fresh tips per transfer (default); confirm plate sealing between stages. |
| Run paused | Vision flagged an exception | Re-seat as prompted; resume. |
When not to automate this
For very low sample numbers, manual kits may be more economical. This protocol is for research/throughput use — clinical diagnostic deployment requires your own validation and regulatory clearance.
Verification: this protocol is simulation-verified — confirmed to run in the simulator but not yet run on hardware by RoR. Validate before using with critical samples.