Introduction to the Workshop
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Abstracts

Mapping

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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.

Large-Scale Oligosynthesis in a Multichannel Format

L.E. Sindelar and J.M. Jaklevic
Human Genome Center and Engineering Division, Lawrence Berkeley Laboratory, University of California, Berkeley, CA 94720

We describe an approach to large-scale parallel oligosynthesis in which a multi-well format is used. The reactions are carried out in open wells using an argon ambient atmosphere to prevent reagent contamination. The controlled-pore glass beads which form the substrate for synthesis are held in individual wells with high-density polyethylene filter bottoms through which reagents are drawn into a vacuum manifold. The synthesis is carried out using direct reagent dispensing into the individual reaction wells. A computer controls the sequence in which reagents are dispensed and the timing of the periodic vacuum pulses required to synthesize the desired sequence.

Experiments to date have demonstrated the viability of the approach for a variety of test sequences. Results obtained with HPLC analysis demonstrate coupling efficiencies as high as 99.5% under optimized conditions. Use of the oligomers for DNA sequencing templates and as PCR primers has been demonstrated in production applications. The current instrument design consists of a series of discrete, 12-channel reaction chambers capable of multiplexing in a 12 x N format where N can be 1 to 8, i.e., 96 wells. A projected time interval for 12 parallel syntheses is 2:30 hours, with 96 syntheses in 3:30 hours. The current 12-channel system is now being used in a full production environment and is easily capable of producing 120 custom oligos per work week. The proposed extension to the larger 12 x N format will provide proportionately greater output. Because of the reduced volume of reagents required in the open well format, significant cost savings are projected.