An Automated Sample Preparation System for Large-Scale DNA Sequencing

Recent advances in DNA sequencing technologies, both in the form of high lane-density gels and automated capillary systems, will lead to an increased requirement for sample preparation systems that operate at low cost and high throughput. As part of the development of a fully automated sequencing system, we have developed an automated subsystem capable of producing 10,000 sequence-ready ssDNA templates per day from libraries of M13 plaques at a cost of $0.29 per sample. This Front End has been in high throughput operation since June, 1997 and has produced > 400,000 high-quality DNA templates.

Improvements in electrophoresis technology have been announced recently by various companies (Molecular Dynamics, Perkin-Elmer/ABI, Beckman Coulter). In particular, the 96-sample capillary electrophoresis instruments being released promise increased throughput and decreased cost for DNA sequencing. Despite improved separation time, automation, and sample tracking compared to slab gels, capillary technologies have not gained acceptance in the past because of difficulties in resolving long DNA strands. Recent developments in sequencing reaction purification (Ruiz-Martine et al. 1998) and their effect on read length have made it more likely that capillary electrophoresis technologies will supplant slab gel-based sequencing instruments in the near future.

http://www.genome.org/cgi/content/full/9/5/457