DNA Sequencing in Noncovalently Coated Capillaries Using Low Viscosity Polymer Solutions

Capillary electrophoresis (CE) has become an alternative to slab-gel electrophoresis for DNA separations due to its many advantages such as speed, increased separation efficiency, requires minute amount of sample, and automation of sample loading (1 ) Currently, high-throughput DNA sequencing is performed exclusively by slab-gel electrophoresis coupled to fluorescence detection systems. However, slab-gel electrophoresis involves labor-intensive steps such as gel pouring, sample loading, and long electrophoretic run times. These disadvantages reduce the overall throughput efficiency of a slab-gel system making it less than ideal. One of the main goals of the Human Genome and other large scale sequencing projects is to increase the throughput rate with a commensurate reduction in the cost-per-base sequenced. A limitation of CE has been that only one capillary could be run and monitored at a time, so that the total experimental throughput is no better than with slower slab-gel system with multiple lanes. However, the recent introduction of the multiple capillary instruments, such as the ABI Prism 3700 DNA Analyzer (2 ) and the MegaBACE (3 ), may have the potential to meet the high throughput demand.