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.

Design of a Parallel Array Oligonucleotide Synthesizer

Thomas Brennan, Scott Hunicke-Smith, Deval Lashkari, and Ronald Davis
Stanford DNA Sequencing and Technology Center, Departments of Genetics and Biochemistry, Stanford University, Stanford, CA 94305

An automated oligonucleotide synthesizer has been developed which can simultaneously and rapidly synthesize up to 96 different oligonucleotides in a standard 96-well format. The machine is capable of both mixed length and mixed scale synthesis, and can accommodate all of the standard internal base and 5'-labeled modifications. Conventional phosphoramidite chemistry is used. The standard synthesis scale is 20 nmol. The appropriate reagents are delivered by banks of valves into the individual wells containing the growing oligonucleotide chain which is bound to a solid support. Each well has a filter bottom to enable the removal of spent reagents. On-line trityl analysis is employed to monitor the coupling efficiency in each well, which is typically > 98%. Oligonucleotides up to 90-mers have been successfully prepared.

With the development of this machine, it has now become practical and cost effective to synthesize thousands to tens of thousands of "custom" oligonucleotides for directed primer walking strategies, as well as PCR mapping, gene and vector construction projects.

This work has been supported by DOE DEFG0393ER6155 and NIH HG00205.