qPCR.md

Research Use Only (RUO). Not for use in diagnostic procedures or patient care.

qPCR for gene expression (example template)

Sources: DOI:xx.xxxx/xxxx; Preprint:

Equipment

• qPCR thermocycler
• Calibrated micropipettes (P10, P200)

Reagents (example)

• SYBR Green Master Mix — 1x final
• Forward primer (10 µM); Reverse primer (10 µM)
• cDNA template

Steps

1. Thaw reagents on ice (≈10 min). Mix by gentle inversion.
2. Prepare master mix (per 20 µL reaction): 10 µL SYBR mix; 0.8 µL fwd primer; 0.8 µL rev primer; x µL template; nuclease-free water to 20 µL.
3. Plate reactions; seal; brief spin.
4. Thermocycler program: 95 °C 2 min; 40 cycles of [95 °C 15 s, 60 °C 60 s]; melt curve.

Safety notes

• Wear PPE; handle reagents per SDS.

Bill of Materials (example)

• SYBR mix (generic), Alt1: Vendor A cat#..., Alt2: Vendor B cat#...