Microchip device for cell lysis, multiplex PCR amplification, and electrophoretic sizing

Anal Chem. 1998 Jan 1;70(1):158-62. doi: 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 lambda 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.

MeSH terms

  • DNA, Bacterial / biosynthesis
  • DNA, Bacterial / genetics
  • Electrophoresis, Polyacrylamide Gel / instrumentation*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Microcomputers*
  • Polymerase Chain Reaction / instrumentation*

Substances

  • DNA, Bacterial