Microchip Device for Cell Lysis, Multiplex PCR Amplification, and Electrophoretic Sizing
- 1 January 1998
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 70 (1) , 158-162
- https://doi.org/10.1021/ac970642d
Abstract
The steps of cell lysis, multiplex PCR amplification, and electrophoretic analysis are executed sequentially on a monolithic microchip device. The entire microchip is thermally cycled to lyse cells and to amplify DNA, and the products are then analyzed using a sieving medium for size separation and an intercalating dye for fluorescence detection. Using a standard PCR protocol, a 500-base pair (bp) region of bacteriophage λ DNA and 154-, 264-, 346-, 410-, and 550-bp regions of E. coli genomic and plasmid DNAs are amplified. The electrophoretic analysis of the products is executed in <3 min following amplification using hydroxyethyl cellulose or poly(dimethylacrylamide) sieving gels. Product sizing is demonstrated by proportioning the amplified product with a DNA sizing ladder.Keywords
This publication has 35 references indexed in Scilit:
- Microchip Separations of Neutral Species via Micellar Electrokinetic Capillary ChromatographyAnalytical Chemistry, 1995
- Continuous Sample Pretreatment Using a Free-Flow Electrophoresis Device Integrated onto a Silicon ChipAnalytical Chemistry, 1994
- Open Channel Electrochromatography on a MicrochipAnalytical Chemistry, 1994
- High-Speed Separations on a MicrochipAnalytical Chemistry, 1994
- Effects of Injection Schemes and Column Geometry on the Performance of Microchip Electrophoresis DevicesAnalytical Chemistry, 1994
- Synchronized cyclic capillary electrophores—a novel approach to ion separations in solutionJournal of High Resolution Chromatography, 1993
- Micromachining a Miniaturized Capillary Electrophoresis-Based Chemical Analysis System on a ChipScience, 1993
- Planar glass chips for capillary electrophoresis: repetitive sample injection, quantitation, and separation efficiencyAnalytical Chemistry, 1993
- Capillary electrophoresis and sample injection systems integrated on a planar glass chipAnalytical Chemistry, 1992
- Planar chips technology for miniaturization and integration of separation techniques into monitoring systemsJournal of Chromatography A, 1992