title: Increasing PCR sensitivity by removal of polymerase inhibitors in environmental samples by using dielectrophoresis.
abstract: Dielectrophoresis (DEP) is a powerful tool to manipulate cells and molecules in microfluidic chips. However, few practical applications using DEP exist. An immediate practical application of a carbon-electrode DEP system, in removing PCR inhibitors from a sample, is reported in this work. We use a high throughput carbon-electrode DEP system to trap yeast cells from a natural sample (fermented grape must) and then in situ remove contaminants that interfere with PCR analysis. Retrieval of this enriched and purified yeast population from the DEP system then allows for a significant increase of sensitivity during PCR analysis. Furthermore, the fact that DEP can discriminate between viable and non-viable cells minimizes the number of false positives commonly obtained when using PCR alone. Experimental results provide clear evidence that the carbon-electrode DEP-based sample preparation step can readily and effectively clean environmental samples from natural contaminants and improve PCR sensitivity.
authors: Institute for Bioengineering of Catalonia (IBEC), Cellular Biotechnology, Baldiri Reixac 15-21, 08028 Barcelona, Spain.
authors: Maria Del Carmen
authors: Mdel C
authors: Jaramillo
authors: Rodrigo
authors: R
authors: Martínez-Duarte
authors: Mario
authors: M
authors: Hüttener
authors: Philippe
authors: P
authors: Renaud
authors: Eduard
authors: E
authors: Torrents
authors: Antonio
authors: A
authors: Juárez
mesh_terms: D002469:Cell Separation / Q000295:instrumentation*; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification*; D004586:Electrophoresis / Q000295:instrumentation*; D004784:Environmental Monitoring / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000295:instrumentation
chemical_list: D004259:DNA-Directed DNA Polymerase