G.M. Yershov, A.I. Belgovsky, L.D. Drobyshev, V.N. Sushkov, N.V. Mologina, D. Guschin, J. Steele, A. Gemmel, A. Zaslavsky, D. Naylor,[1] and A.D. Mirzabekov
Argonne National Laboratory (U.S.A.) and Engelhardt Institute of Molecular Biology (Moscow)-Joint Human Genome Program.
The manufacture of matrices with gel-immobilized oligonucleotides (sequencing microchips) to be used in SHOM (sequencing by hybridization with oligonucleotide microchips) includes three separate stages. The first stage is shaping the desired topology of the gel micromatrix by mechanical scribing, laser ablation, or photopolymerization. The second stage is to load microvolumes of oligonucleotide solutions onto the matrix of the gel "cells", and the third is to immobilize the oligonucleotides within the cells. The production rate of such matrices is limited mainly by the step of loading microvolumes.
We have designed two production robots for rapid microdispensing of the oligonucleotides which have to be immobilized onto the cells of microchips. The first robot was established in Moscow and successfully tested. It filled five cells per minute using one pin and 80 cells per minute using 16 pins. The other robot, developed at Argonne National Laboratory under the joint project, has higher productivity and accuracy. It can fill approximately 160 cells per minute using 16 pins and 640 cells per minute using 64 pins. Productivity can be increased by producing many similar microchips in parallel. The development of a fully automated line is well under way. Major features of this work will be highlighted in a poster session at the conference.
Using three-dimensional gels as carriers of oligonucleotides, microchip manufacturing technology and hardware have been developed. This approach can also be successful (with or without chemical modification of various gels) for the detection of specific chemical interactions of water-soluble bioorganic compounds. This subject will also be discussed in the poster session.
* Work supported by the U.S. Department of Energy Office of Health and Environmental Research, under contract No. W-31-109-ENG-38; by the Affymetrix, Inc., U.S.A.; and the Russian Human Genome Program.
[1] University of Illinois at Chicago (U.S.A.)
The submitted manuscript has been authorized by a contractor of the U.S. Government under contract No. W-31-109-ENG-38. Accordingly, the U.S. Government retains a nonexclusive, royalty-free license to publish or reproduce the published form of this contribution, or allow others to do so, for U.S. Government purposes.