A. Mirzabekov, G. Yershov, V. Barsky, E. Timofeev, D. Guschin, V. Shick, S. Dubiley, D. Pobedimskaya, and D. Prudnikov
Argonne National Laboratory (U.S.A.) and Engelhardt Institute of Molecular Biology (Moscow)-Joint Human Genome Program.
The development of biological microchips will allow us to collect and analyze significant amounts of biological information in comparatively simple experiments. Oligonucleotide microchips have been manufactured and applied for analysis of DNA sequences. The microchips (40x40x20 mm or larger size) consist of gel elements with immobilized oligonucleotides fixed on a glass surface. A robot was constructed for large-scale microchip production, in which it applies activated oligonucleotide solutions (1-50 nl) to gel elements. A simple method was also developed for manual microchip manufacturing. The hybridization of fluorescently labeled DNA with a microchip is monitored in real time at several wavelengths, and with a temperature gradient by means of a specially devised fluorescent microscope coupled with a CCD camera. Alternatively, the hybridized unlabeled DNA can be stained directly on the microchip with a fluorescent dye. Contiguous stacking hybridization has been developed to raise the sequencing efficiency of, for example, an 8-mer microchip to that of a 13-mer microchip. The equipment and the method developed were applied for analysis of DNA sequences and as diagnostics for genetic diseases. The development of SHOM for mapping, as well as partial and complete sequencing of DNA will be presented in posters.
Our technology allows us to manufacture microchips bearing different immobilized compounds of biological interest, such as various oligonucleotides, peptides, DNA, RNA, proteins, antibodies, and low molecular weight ligands. Potential applications for such biological microchips will be discussed.
*Work supported in part by the U.S. Department of Energy, Office of Health and Environmental Research, under Contract No. W-31-109-ENG-38 and Russian Human Genome Program.
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.