A novel thermocycling machine based on a microcapillary equipped with bidirectional pressure-driven flow and in situ optical position sensors is described. A 1-microL droplet of reaction mixture moves between three heat zones in a 1-mm-i.d., oil-filled capillary using a multielement scattered light detector and active feedback. Dwell times and accelerations can be adjusted independently. As a demonstration of the device, 30 cycles of a 500-base pair product were performed in 23 min with 78% amplification efficiency. This result compares well with previous high-speed thermocyclers. Theoretically, the arrangement can approach a time of 2.5 min for 30 cycle amplifications of a 500-base pair product.

An experimental study is performed on the internal cooling of a rotating serpentine flow passage of square cross section with throughflow. The test section is not proceeded by a hydrodynamic calming region, i.e., a leading arm, and is rotated at low Rossby numbers. The local heat transfer coefficients along the flow passage, including the leading wall, trailing wall, and sidewalls, are determined together with the circumferentially averaged values. The Reynolds, Rossby, and rotating Rayleigh numbers are varied to determine their effects on heat transfer performance. It is disclosed that heat transfer augmentation is significant at all sharp turns due to the presence of strong secondary flow. The rotational effect is very obvious and complicated in the local heat transfer performance but it is very minor on the average heat transfer performance. The throughflow rate plays an important role on the heat transfer performance. The results may serve as a baseline for comparison with the results from a model with a leading arm to determine the effects of a hydrodynamic calming section on the heat transfer performance of a rotating serpentine flow passage.

The performance of a novel thermal cycler has been characterized in a 30-cycle PCR. The device consists of a microcapillary equipped with bidirectional pressure-driven flow and in situ optical position sensors. A 1-µL droplet of reaction mixture moves between three heat zones in a 1-mm i.d., oilfilled capillary using a multi-element scattered light detector and active feedback. The design permits time and number of cycles to be changed without hardware modification, unlike other flow-in-capillary PCR systems. Temperature optimization has been performed on the three PCR heat steps. The optimal denaturation temperature is 94°C-96°C, which is identical to commercial machines. The optimal extension temperature of 62°C-66°C is lower than reported for Taq DNA polymerase (70°C-80°C) because of the high enzyme concentration and/or the absence of detergent in the PCR mixture. The optimal annealing temperature seems to be the same as the optimal extension temperature. This is because extension occurs when the sample is inside of the annealing heat zone. Annealing takes place as the sample travels between heat zones. Device speed (23 min for 30 cycles without time optimization) is competitive with other rapid PCR designs for efficiencies comparable to a commercial machine.

No abstract

Experiments are performed to determine the local heat transfer performance in a rotating serpentine passage with rib-roughened surfaces. The ribs are placed on the trailing and leading walls in a corresponding posited arrangement with an angle of attack of 90 deg. The rib height-to-hydraulic diameter ratio, e/Dh, is 0.0787 and the rib pitch-to-height ratio, s/e, is 11. The throughflow Reynolds number is varied, typically at 23,000, 47,000, and 70,000 in the passage both at rest and in rotation. In the rotation cases, the rotation number is varied from 0.023 to 0.0594. Results for the rib-roughened serpentine passages are compared with those of smooth ones in the literature. Comparison is also made on results for the rib-roughened passages between the stationary and rotating cases. It is disclosed that a significant enhancement is achieved in the heat transfer in both the stationary and rotating cases resulting from an installation of the ribs. Both the rotation and Rayleigh numbers play important roles in the heat transfer performance on both the trailing and leading walls. Although the Reynolds number strongly influences the Nusselt numbers in the rib-roughened passage of both the stationary and rotating cases, Nuo and Nu, respectively, it has little effect on their ratio Nu/Nuo.