DOUBLER SEQUENCING IN MOLECULAR DIAGNOSIS OF HEREDITARY-DISEASES

We describe doubler sequencing of human genomic PCR products using two differently labeled primers in a single reaction and analysis on two automated DNA sequencing devices. Feasibility of the methodology is de monstrated by isothermal and cycle sequencing for two different PCR pr oducts and by cycle sequencing on both strands of a single product. It was applied to analyze mutations in patient DNAs in routine sample sc reening. Because it has the advantage of increased throughput and cost reduction while retaining its accuracy and reading length, we found t hat doubler sequencing is an attractive option for molecular diagnosis of hereditary diseases. This approach would be even more beneficial i f it used DNA sequencing devices with several lasers in a single instr ument.