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Rapid PCR* Using Active Tube Temperature Control On Thermo's PCR Machines

Thermo Electron Corporation
Solutions Note

The time taken to complete a standard PCR protocol is becoming increasingly important for researchers today. Whilst many manufacturers of thermal cyclers focus on pushing the limits of ramp rates, in this article we highlight how Thermo's approach of focussing on achieving accurate sample temperatures can yield PCR results in less than 1 hour.

Abstract

The Active Tube Control feature of Thermo's thermal cyclers measures the actual temperature of the reaction mix during PCR. This enables reduced dwell time PCR programmes to be run. A one second dwell time PCR programme was successfully run on Thermo's MBS and Px2 thermal cyclers. Yield of PCR product was comparable to that obtained from a standard PCR programme with a 30 second dwell time, but total run time was reduced by up to 50%.

Introduction

A standard 3 temperature PCR programme can take in excess of 1 hour 30 minutes to complete. For modern researchers it is desirable to reduce these experimental times to speed up laboratory workflow. The use of Active Tube Temperature Control provides accurate monitoring and adjustment of in-sample reaction conditions, significantly reducing reaction dwell times and, therefore, experimental times without affecting PCR product yield.

Thermo thermal cyclers can use up to five control modes for monitoring temperature during PCR reactions. Block Control is the sole control method used on some thermal cyclers. As the name implies, the block temperature is monitored during the PCR reaction, irrespective of the in-sample temperatures achieved. This approach is adequate for prolonged static incubations. However, if Block Control is used during short incubation periods, such as during a PCR reaction, the reaction mix only achieves the set temperature for
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