Thermal control

Efficient performance of quantitative PCR requires rapid, precise, thermal control.

30 cycles of PCR have been demonstrated in less than 10 minutes. Rapid cycling provides several benefits, including, reduced time to result, increased system throughput and improved reaction specificity. In practice however, engineering trade-offs between ease of use, temperature uniformity, and speed, mean that reaction times are typically more than 25 minutes.

Thermal non-uniformity during temperature cycling contributes to variability in PCR and, unfortunately, some thermocyclers do not meet the specifications claimed by manufacturers. Increasing the speed of thermal cycling generally reduces thermal uniformity, and can reduce the precision of real-time PCR.

The temperature uniformity also has a direct impact on the ability to discriminate different PCR products by performing melting point analysis. In addition to uniformity, the resolution with which instruments are able to control temperature is a factor which affects their performance when performing high resolution melting analyses.

Therefore speed, precision and uniformity of thermal control are important performance characteristics of real-time PCR instruments.