Microfluidic genetic analysis

Kamei T, Toriello NM, Lagally ET et al (2005) Microfluidic Genetic Analysis with an Integrated a-Si H Detector. Biomed Microdevices 7 147-152... 

Fig. 16 Microfluidic genetic analysis (MGA) system, (a) Dyes are placed in the channels for visualization Scale bar. 10 mm). Domains for DNA extraction yellow), PCR amplification red), injection green), and separation blue) are connected through a network of channels and vias. SPE reservoirs are labeled for sample inlet ST), sidearm ( 4), and extraction of waste (EW). Injection reservoirs are labeled for the PCR reservoir PR), marker reservoir (MR), and sample waste (5W). Electrophoresis reservoirs are labeled for the buffer reservoir (BR) and buffer waste (BW). Additional domains patterned onto the device included the temperature reference TR) chamber and fluorescence alignment (FA) channel. The flow control region is outlined by a dashed box. Device dimensions are 30.0 x 63.5 mm with a total solution volume < 10 pL Scale bar. 10 mm), (b) Flow control region. Valves are shown as open rectangles. VI separates the SPE and PCR domains. V2 and V5 are inlet valves for the pumping injection, V3 is the diaphragm valve, and V4 is an outlet valve, (c) Device loaded into the manifold, (d) Intersection between SI and SA inlet channels, with the EW channel tapering to increase flow resistance Scale bar. 1 mm).

Easley, C. J., Karlinsey, J. M., Bienvenue, J. M., Legendre, L. A., Roper, M. G, Feldman, S. H., Hughes, M. A., et al., A fuUy integrated microfluidic genetic analysis system with sample-in-answer-out capability, Proceedings of the National Academy of Sciences of the United States of America 103, 19272-19277, 2006. 

Easley, C.J. et al., A fully integrated microfluidic genetic analysis system with sample-in-answer-out capability. Proc. Natl. Acad. Sci. USA, 103, 19272, 2006. 

Integrated Microdevices for Biological Applications, Fig. 3 Images of the microfluidic genetic analysis device. Domains for DNA extraction (d), PCR amplification (e), injection, and separation (f) are connected through a network of channels and vias (Reprinted with permission from Easley et al. [6])... 

Easley et al. [6] reported a microfluidic genetic analysis system capable of accepting whole blood as a crude biological sample with the endpoint generation of a genetic profile (Fig. 3). 

Huang et al. [176] described an integrated microfluidic chip (of PDMS and soda-lime glass) capable of performing DNA/RNA amplification, electroki-netic sample injection and separation, and online optical detection in an automatic mode. The authors tested its functionality for bacterial DNA of Streptococcus pneumoniae and RNA of dengue-2 vims. The NCE developed represented a crucial contribution to the fields of molecular biology, genetic analysis, infectious disease detection, and other biomedical applications. 

The microfluidic chip uniquely facilitates high-throughput genetic analysis. For instance, 12 different samples were analyzed in parallel by capillary array electrophoresis (CAE) see Figure 9.20. The two-color method was used for... 

Keywords Continuous flow PCR DNA analysis DNA microarrays Genetic analysis Integrated microsystems Microcapillary electrophoresis Microfluidics Micro-PCR devices Solid-phase extraction... 

Finally, on-chip DNA quantitation will undoubtedly be an important feature of microfluidic sample processing for a number of different applications where the amount of input DNA for downstream genetic analysis is crucial. As discussed more extensively in Chapter 37, quantitative PCR (qPCR) methods in microfluidic systems have not been demonstrated to date. However, there are a number of publications describing real-time PCR in microdevices and it is likely only a matter of time before... 

Liu, R. H., and Grodzinski, R, Development of integrated microfluidic system for genetic analysis, J. Microlith. Microfab. Microsyst., 2, 340, 2003. 

By combining facile electrical measurements with controllable physical and chemical stimuli at the micron level, microfluidic devices are developing into a versatile platform for cell-based biosensors, drug discovery, genetic analysis, and diagnostics. 

Current developments are not only aimed at integration but also at parallel genetic analysis as well. Parallel genetic analysis allows detection of different DNA samples simultaneously. A microfluidic device integrated with multichamber PCR and multichannel CE for parallel genetic analysis was developed (Figure 10.4) [56]. The microdevice consists of three functional units temperature control, multiple PCR (four chambers PCR), and multiple channel separation (four separation channels, each channel cormected to a PCR chamber). A separation channel is... 

Many advantages have been reported with the use of microfluidic reactors in biocatalytic reactions. These advantages can potentially enable the rapid evaluation of different reaction conditions, overcoming the time constraints associated with biocatalytic process development. Biocatalysis by enzymatic microreactors have been widely reported. Enzymatic microreactors are classified based on their applications such as microreactors for enzymatic diagnosis and genetic analysis, for enzyme-linked immunoassays, and for analysis of proteins. Microreactors for genetic analysis have been widely exploited, while integration of microfluidics... 

Thompson AM, Paguirigan AL, Kreutz JE et al (2014) Microfluidics for single-cell genetic analysis. Lab Chip 14 3135-3142... 

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