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

es cover only approximately 60% of the bottom surface of the block; this design creates a temperature gradient from the center to the edges of the block. Moreover, to ramp temperatures up and down quickly, use of the Peltier modules requires that the blocks mass be limited.

Maintaining a consistent temperature across a thermal block is easier when the exposed surface area is kept to a minimum, thereby limiting heat loss due to convection. Each hot block of the RoboCycler temperature cycler maintains its programmed temperature throughout the course of a PCR amplification experiment. A robotic arm moves PCR samples from one preset hot block to another at user-specified times. This eliminates the need to ramp blocks up and down in temperature and permits the use of blocks with minimal surface area. Consequently, RoboCycler temperature cyclers maintain a more consistent temperature throughout all wells of the block and, hence, provide improved PCR results.

Fluid Temperature Profiles: RoboCycler 96 vs. MJ Research PTC-100

Figure 1

To compare well-to-well temperature consistency and sample ramping rate, fluid temperature profiles were determined during thermal cycling using both a RoboCycler 96 temperature cycler fitted with Stratagenes Hot Top Assembly and an MJ Research PTC-100 with heated lid enabled. In each case, fluid temperature measurements were taken in six tubes positioned in the four corner wells and two center wells as shown in Figure 1. Similar testing parameters were used for each thermal cycler. T-type thermocouple temperature probes were inserted into six, 200-l thin-walled tubes containing 50 l of 1X Taq DNA polymerase buffer and cycled through three cycles of 95C for 1 minute, 55C for 2 minutes, and 72C for 1 minute. The temp
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