Input
Primer Sequences
Only A, C, G, T allowed. Spaces and line breaks will be ignored.
Only A, C, G, T allowed. Spaces and line breaks will be ignored.
PCR Settings (Optional)
These values are used for simple Tm estimation and an approximate PCR mix. You can leave defaults if you’re not sure.
Results
Summary
No analysis yet
Primer Metrics
| Metric | Forward | Reverse |
|---|---|---|
| Length (nt) | – | – |
| GC content (%) | – | – |
| Tm (°C) | – | – |
| 3'-end GC clamp | – | – |
| Basic validity | – | – |
Secondary Structure & Dimers
Self-dimers
Primer–primer dimers
Other flags
Suggested Annealing Temperature
Recommended Ta: –
Gradient suggestion: –
Rule-of-thumb: Ta ≈ Tmlower − 3–5 °C. This is a simplified estimate for quick planning.
PCR Mix (Suggested)
Based on your reaction volume and primer concentrations. Always adjust for your specific polymerase / protocol.
| Component | Stock | Final | Volume (µL) |
|---|