title: "Experiment log: qPCR standard curve — April sample run" project: research-lab-template tags:

• experiment
• qpcr
• qc date: 2026-04-07

qPCR standard curve — April sample run

Operator: Template user | Instrument: qPCR-01 | Kit lot: DEMO-LOT-001

Objective

Establish a six-point standard curve for housekeeping and target amplicons before running unknowns from the cytokine panel pilot.

Setup

100 ng/µL RNA, 1:5 serial dilutions (six tubes); triplicate 10 µL reactions with ROX per manufacturer.

Observations

Standards show single melting peaks (62°C anneal); NTCs undetermined at 40 cycles. Layout: standards cols 1–3; NTC col 4; unknowns from col 5 next run.

Data (summary)

Cq: 1e0 mean 18.4 (SD 0.12); 1e-5 mean 35.1 (SD 0.21). Slope efficiency ≈ 98% (target band 90–110%).

Issues

• Well A7 showed high fluorescence before cycle 1—excluded as meniscus bubble; repeat if unknowns border that well.

Next steps

Import Cq into the analysis notebook (link path in a child note); run unknowns on the same master-mix lot; append protocol with 30s denature (beats 10s in our test).