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Effects of Thermal Cycler Well-To-Well Temperature Uniformity on PCR Results


Better temperature uniformity allows for better control of PCR reactions

Michelle Cayouette Jane Moores Jason Capodanno Michael Rahilly
Stratagene

Stratagenes RoboCycler temperature cycler # produces consistent PCR results in every well. The RoboCycler systems unique design uses thermal blocks and heating techniques that are optimized for block temperature uniformity. In this study, fluid temperature profiles and PCR results were determined using both a RoboCycler 96 temperature cycler and a typical Peltier single-block thermal cycler, the MJ Research PTC-100. The RoboCycler system showed better fluid temperature well-to-well uniformity, faster fluid temperature ramping rates, and more consistent PCR-product banding patterns.

When performing PCR amplifications, reaction components and PCR parameters are not the only criteria that affect the quality of results; differences may also exist in the ability of thermal cyclers to maintain a consistent temperature both within and between experiments. If the well-to-well temperature across the thermal block is not uniform, it can have a significant impact on the PCR. This is an especially important consideration for researchers performing quantitative or diagnostic PCR assays using multiple samples.

The four-block design common to all members of Stratagenes RoboCycler line of thermal cyclers provides a higher degree of well-to-well uniformity than the single-block design of many other thermal cyclers. Each of the three RoboCycler hot blocks is heated using a microprocessor-controlled resistive element heater, which covers 95% of the bottom surface of each block. Other thermal cyclers, such as the MJ Research thermal cyclers, use Peltier thermal electric modules attached to the bottom of a single block for heating and cooling. However, the Peltier modul
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