Introduction to the Workshop
URLs Provided by Attendees

Abstracts

Mapping

Informatics

Sequencing

Instrumentation

Ethical, Legal, and Social Issues

Infrastructure

The electronic form of this document may be cited in the following style:
Human Genome Program, U.S. Department of Energy, DOE Human Genome Program Contractor-Grantee Workshop IV, 1994.

Abstracts scanned from text submitted for November 1994 DOE Human Genome Program Contractor-Grantee Workshop. Inaccuracies have not been corrected.

A Rapid, Microtiter-Plate, Robotics-Compatible, Multi-Station Thermal Cycler

A.D.A. Hansen, J.M. Jaklevic, V.M. Stevko, W.F. Kimmerly, et al.
Human Genome Center and Engineering Division, Lawrence Berkeley Laboratory, University of California, Berkeley, CA 94720

We have developed a thermal cycler based on the circulation of temperature-controlled water directly to the underside of thin-walled polycarbonate microtiter plates. The water is rapidly switched from a set of reservoirs. The plate wells are loaded with typically 15-20 µl of reagent mix for the PCR process. Heat transfer through the thin polycarbonate is sufficiently rapid that the contents reach thermal equilibrium with the water in less than 15 seconds, although unexpected improvements in performance are observed in non-equilibrium runs in which the temperature switching is more rapid. Complete PCR amplification runs of 40 three-step cycles have been performed in as little as 14.5 minutes, with the results showing substantially enhanced specificity compared to conventional technology requiring run times in excess of 100 minutes. 96-, 192- and 384-well plates can be used. The plate clamping station includes a heated lid, eliminating the need for mineral oil overlay of the reactants, and was specifically designed to be accessible for robotic loading and unloading of the plates. The present apparatus has three plate stations, fed from common reservoirs but operating with independent switching cycles. The high overall throughput is required to meet the needs of the LBL Human Genome Center effort. The enhanced performance and specificity observed in some cases may allow for extension of PCR methods in mapping and sequencing strategies. Experience developed in using the system in a production environment will be described together with future design modifications.