Chip PCR. II. Investigation of different PCR amplification systems in microbabricated silicon-glass chips

Nucleic Acids Res. 1996 Jan 15;24(2):380-5. doi: 10.1093/nar/24.2.380.

Abstract

We examined PCR in silicon dioxide-coated silicon-glass chips (12 microl in volume with a surface to volume ratio of approximately 17.5 mm(2)/microl) using two PCR reagent systems: (i) the conventional reagent system using Taq DNA polymerase; (ii) the hot-start reagent system based on a mixture of TaqStart antibody and Taq DNA polymerase. Quantitative results obtained from capillary electrophoresis for the expected amplification products showed that amplification in microchips was reproducible (between batch coefficient of variation 7.71%) and provided excellent yields. We also used the chip for PCR directly from isolated intact human lymphocytes. The amplification results were comparable with those obtained using extracted human genomic DNA. This investigation is fundamental to the integration of sample preparation, polynucleotide amplification and amplicate detection on a microchip.

Publication types

  • Comparative Study

MeSH terms

  • Antibodies
  • Base Sequence
  • Campylobacter jejuni
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics
  • DNA / analysis*
  • DNA / blood
  • DNA, Bacterial / analysis
  • DNA-Directed DNA Polymerase
  • Electrophoresis, Capillary
  • Glass
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction / instrumentation
  • Polymerase Chain Reaction / methods*
  • Silicon
  • Silicon Dioxide
  • Taq Polymerase

Substances

  • Antibodies
  • CFTR protein, human
  • DNA, Bacterial
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Silicon Dioxide
  • DNA
  • Taq Polymerase
  • DNA-Directed DNA Polymerase
  • Silicon