Flow-Through PCR

Alternative methods have been reported to achieve temperature variations needed for PCR. 

Similar methods of flow-through PCR chips were also reported [212,907,948], Temperature analysis of continuous flow PCR has been carried out on both a glass-Si and glass-glass chip [949]. 

In another report, a PDMS rotary device (12 nL) was used for PCR both spatially (moving over three temperature zones) or temporally (at three temperatures with rotary mixing). PCR of a P-actin gene (123 bp) and X phage DNA (199 bp) were demonstrated. Real-time PCR was also performed as detected by an intercalating dye (Sybr Green I) [357]. 

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Applications to Nucleic Acids Analysis 9.2.1.5 Flow-Through PCR... 

A sophisticated integrated flow-through PCR chip was fabricated (see Figure 9.11). Three reaction chambers, connection channels, thin-film resistive heaters, temperature sensors, and optical detectors are fabricated on a Si wafer. A thin... 

Schneegass, I., Brautigam, R., Kohler, J.M., Miniaturized flow-through PCR with different template types in a silicon chip thermocycler Labchip 2001, 1, 42-49. 

Figure 14 (A) Schematic of a chip for flow-through PCR. Three well-defined zones are kept at 95, 77, and 60°C by means of thermostated copper blocks. The sample is hydrostatically pumped through a single channel etched into the glass chip. The channel passing through the three temperature zones defines the thermal cycling process. (B) Layout of the device used in the study. The device has three inlets on one side of the device and an outlet on the opposite side. Only two inlets were used, one carrying the sample, the other bringing a constant buffer flow. The chip incorporates 20 identical cycles, except for the first one, which includes a threefold increase in DNA melting time. (Reprinted with permission from Ref. 98.)...

Fig. 2 Layout of a PCR chip reported by Manz et al. (A) Schematic of a chip for flow-through PCR. (B) Layout of the device used in the Manz study. (From Ref. f)...

FIGURE 43.3 Schematic views of the flow-through PCR device layout and model. The device on the right was used in the experiments described and had 50-mm wide channels separated by 250 mm. The device on the left had 50-mm wide channels separated by 50 nun and was not nsed in the research presented, however, it illustrates the versatility of the flow-through design. (Figure adapted and reproduced from Hashimoto, M., et al., Lab Chip, 2004, 4, 638-645. Copyright 2004. With permission from Royal Society of Chemistry.)... 

The principle of flow-through PCR can further be miniaturized and optimized with respect to heat transfer conditions by realizing flow-through chip thermocyclers [11, 12]. These are devices... 

The evaporation of solvent can be avoided completely if closed microchannels are used for a micro-PCR with moving of samples between different temperature zones. The simplest variant consists of a flow-through PCR realized in a tube, which is conducted repeatedly through the three temperature zones. There are two principal geometries for the arrangement of tubes. In one variant the tube is coiled in one plane like a spiral (Fig. 6a). This geometry can be... 

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