Protocols
General reaction setup and cycling conditions for primerprobe assays. Assay-specific notes are listed on each product page.
Reaction setup (20 µL)
| 2X qPCR master mix | 10.0 µL |
| Primer–probe mix (20X) | 1.0 µL |
| Template DNA / cDNA | 2.0 µL |
| Nuclease-free water | 7.0 µL |
| Total reaction volume | 20.0 µL |
Cycling program
| Step | Temp | Time | Cycles |
|---|---|---|---|
| Initial denaturation / UDG | 50 °C | 2 min | 1 |
| Polymerase activation | 95 °C | 2 min | 1 |
| Denaturation | 95 °C | 15 s | 40 |
| Annealing / extension (read) | 60 °C | 45 s | 40 |
Handling & storage
- Store primer–probe mixes at −20 °C, protected from light.
- Thaw on ice, vortex briefly and spin down before use.
- Avoid more than 10 freeze–thaw cycles; aliquot for routine use.
- Always include a no-template control and, where supplied, the internal control.
For research use only. Not for use in diagnostic procedures.