CAPILLARY AUTOMATED SUBMICROLITER SAMPLE PREPARATION

University Of Washington, Seattle WA

Investigators

Linked publications & trials

Abstract

The long term objective of this proposal is to automate and improve a key area and current bottleneck of genomic research: the automated handling of submicroliter fluid samples. Specifically, the research proposed in this application is the development of a novel, high-throughput, high-yield, fully automated system capable of performing restriction enzyme digests, PCPs, and sample preparation for DNA sequencing. The immediate goals of this system are to process 5,000 1 mu l samples per day, at a cost per finished sequence base pair of less than 20 cents. This will be accomplished through a 10 fold reduction in reagents and sample volumes, and a 10 fold increase in throughput derived from automation, parallelization, and integration of: (I) prior technology developed by the proposing team, and (2) new technology that will be developed during the course of this project. The system will interface in an automated fashion with microtiter plates, Eppendorf tubes, or capillaries at the front end, and with electrophoretic capillaries, electrophoretic gel loading combs, or microtiter plates at the back end. Other input/output media are also possible. The proposed system uses glass capillary tubes and several novel methods to reduce sample size, automate the handling of small fluid samples, reduce thermal cycling and incubation times, and minimize the amount of disposables used to perform DNA sequencing. All processing steps are performed within these capillaries. This use of capillaries also helps facilitate sample containment and minimize evaporation losses, which are particularly important for the high surface area to volume ratios of small samples. An important feature of the proposed system is that it is consistent with current methods of sample handling. Furthermore, it will reduce costs, increase yield, and increase throughput in deriving DNA sequence. The system described in this proposal will greatly aid the Human Genome Project in meeting its sequencing goals. It will also have a synergistic effect outside of the Human Genome Project, benefiting efforts in clinical testing, medical diagnostics, pharmaceutical development, environmental testing, DNA fingerprinting, and agricultural research.