By Gestin, Goldstein
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Lab. Chip. 3(1), 11N–19N. 5. Selvaganapathy, P. , Carlen, E. , and Mastrangelo, C. H. (2003) Recent progress in microfluidic devices for nucleic acid and antibody assays. Proc. IEEE 91(6), 954–975. 6. , Nielsen, P. , Jeffares, D. , et al. (2004) Direct isolation of poly(A)+ RNA from 4 M guanidine thiocyanate-lysed cell extracts using locked nucleic acid-oligo(T) capture. Nucleic Acids Res. 32(7), e64. Porous Polymer Monoliths 21 7. , and Gilgen, M. (2003) Comparative evaluation of four large-volume RNA extraction kits in the isolation of viral RNA from water samples.
The PDMS (ratio between A and B parts is 10:1) was poured onto the master and baked at 80°C to yield a replica. Finally, the replica was mounted to a precleaned slide to form a device with desired channels. E. 5 μm wide and less than 2 μm long. Culturing the GFP-expressing E. coli in LB medium containing ampicillin ensures the presence of GFP plasmid. To determine the performance of the device, GFP-expressing E. coli cells were loaded into the cell reservoir at the cathode region and treated with a DC electric field.
The time for the replica to be immersed in the acid solution needs to be limited to an hour. The PDMS can absorb water and become opaque. This would interfere with the fluorescence microscopy. 4. Treating the replica and slide with an ozone cleaner is also an effective way to make the surface hydrophilic. The PDMS replica will seal the slide irreversibly after being treated by the ozone cleaner. 5. The channel surface will regain hydrophobicity quickly when exposed to the air. After the PDMS replica is taken out of HCl solution and blown dry, the PDMS replica is immediately sealed to a clean glass slide and phosphate buffer is filled in.