HOT

High Temperature Red Alert

Is your PCR machine suffering from extreme heat when the temperature exceeds 35℃?

Poor cooling is stealing your detection accuracy!

qPCR amplification curves splitting abnormally? Amplification efficiency unstable?

Cq values drifting? Spurious peaks in melting curves?

Don’t Panic

This protection guide will help your instruments survive the summer safely!

3 major strategies to block thermal interference,

keeping your ΔΔCq values rock‑steady!

01. Watch Out for PCR Machine “Heatstroke” in High Temperatures

When temperatures rise above 35℃, the cooling efficiency of PCR instruments drops sharply! Carefully designed experiments can be ruined by poor equipment layout. To protect your lab, watch for these 5 warning signs of PCR “heatstroke”:

This proven golden lab layout will save your data from heatwaves:

● Leave ≥30 cm clearance between the instrument and walls on back/side;

● Keep away from heat sources like centrifuges and refrigerators; surface temp of nearby devices ≤40℃;

● Voltage fluctuation ≤5% (online UPS recommended for stable power);

● Avoid direct airflow from AC (30° angled airflow is ideal);

● Prevent direct sunlight exposure.

02. Experimental Protection

4‑Step Ice Method for Sample Processing:

● Grinding: full coverage with liquid nitrogen;

● Lysis: pre‑chilled lysis buffer + operation on ice;

● Reverse transcription: hot‑start enzyme + pre‑aliquoted mix;

● Loading: cold block operation (≤4℃);

Besides manual protection, you can choose Rocgene’s ArchiPure series automated nucleic acid extractors with matched reagents to shorten assay time and free hands, making extraction easy even in hot weather!

Reference Gene Selection Rules:

● Use caution with genes like GAPDH, whose expression increases under heat stress;

Anti‑Evaporation Tips:

● Volume deviation: prepare master mix first, then aliquot on ice using 8‑channel pipette;

● Edge‑well concentration: double sealing with film + mineral oil (Archimed series uses hollow‑block design with temperature compensation to reduce edge effects).

03. Data Analysis Correction

Amplification Efficiency Correction:

The Archimed series not only features hardware temperature compensation but also supports temperature correction in the ArchimedAnalyzer software. It includes both ΔΔCq and double‑standard‑curve relative quantification methods. The ΔΔCq module allows input of amplification efficiencies for reference and target genes to correct data, making high‑temperature ΔΔCq results more reliable!

Melting Curve Correction:

● Peak width >0.8℃ → re‑optimize annealing temperature;

● Spurious peak height >15% of main peak → discard the data point;

ArchimedAnalyzer software supports melting curve parameter settings to filter out false peaks: