Degenerate oligonucleotide primed-polymerase chain reaction and capillary electrophoretic analysis of human DNA on microchip-based devices.

Random amplification of the human genome using the degenerate oligonucleotide primed–polymerase chain reaction (DOP-PCR) was performed in a silicon– glass chip. An aliquot of the DOP-PCR amplified genomic DNA was then introduced into another sili- con–glass chip for a locus-specific, multiplex PCR of the dystrophin gene exons in order to detect dele- tions causing Duchenne/Becker muscular dystrophy. Amplicons were analyzed by both conventional capil- lary electrophoresis and microchip electrophoresis and results were compared to those obtained using standard non-chip-based PCR assays. Results from mi- crochip electrophoresis were consistent with those from conventional capillary electrophoresis. Whole genome amplification products obtained by DOP-PCR proved to be a suitable template for multiplex PCR as long as amplicon size was <250 bp. Successful detec- tion and resolution of all PCR products from the mul- tiplex PCR clearly shows the feasibility of performing complex PCR assays using microfabricated devices.