DOE Human Genome Program Contractor-Grantee Workshop VIII
February 27-March 2, 2000  Santa Fe, NM

45. MicroArray of Gel Immobilized Compounds on Chip

V. Vasiliskov, A. Stomakhin, B. Strizhkov, S. Tillib, V. Mikhailovich, A. Sobolev, A. Kuhktin, and A. Mirzabekov

Joint Human Genome Program: Biochip Technology Center, Argonne National Laboratory, Argonne, IL 60439 and Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 117984

Micro Array of Gel Immobilized Compounds on a Chip (MAGIChip^TM) are produced by immobilizing oligonucleotides¹, DNA, enzymes, antibodies, and other proteins² on a photopolymerized micromatrix of polyacrylamide gel pads. Recently, a photoco-polymerization technique was introduced for more rapid and inexpensive manufacturing of the microchips containing gel pads from 10´10´5 m to 100´100´20 m and larger in size³.

MAGIChips are efficient for carrying out direct hybridization tests, as well as for oligonucleotide ligation, single-base extension⁴, and PCR amplification of DNA. The fluorescence microscope has been devised for quantitative and real-time monitoring of hybridization, measuring the thermodynamic parameters of DNA duplexes, and measuring kinetics of enzymatic reaction on MAGIChips. On-chip MALDI-TOF mass spec-trometry was successfully tested for polymorphism analysis of DNA⁵. These technologies are demonstrated for identification of microorganisms, detection of their genes, and screening for mutations. Simple equipment and procedures have been developed for monitoring the hybridization of amplified DNA with oligonucleotide microchips and on-chip amplification. This allows us to carry out fast and inexpensive screening of Mycobacterium tuberculosis drug resistant mutations.

Work supported by the Department of Energy, Office of Health and Environmental Research under Contract No. W-31-109-ENG-38; Cooperative Research and Development Agreement No. 970192 between Argonne National Laboratory, Motorola, and Packard Instruments; Defense Advanced Research Project Agency under Interagency Agreement No. AO-E428; and the Russian Foundation of Fundamental Research under Grant 96-04-49858.

References

1. Yershov, G., Barsky, V., Belgovskiy, A., Kirillov, Eu., Kreindlin, E., Ivanov, I., Parinov, S., Guschin, D., Drobishev, Dubiley, S. & Mirzabekov, A. (1996) Proc. Natl. Acad. Sci. USA 93, 4913-4918.

2. Arenkov, P., Kuhktin, A., Gemmell, A., Chupeeva, V., & Mirzabekov, A. (2000) Anal. Biochem. (in press).

3. A. Vasiliskov, V., Timofeev E., Surzhikov S., Drobyshev, A., Shick, V., & Mirzabekov, A. BioTechnique 27, 592-606

4. Stomakhin, A, Vasiliskov, V., Timofeev, E., Schulga, D., Cotter, R., & Mirzabekov, A. (2000) Nucleic Acids Res. (in press).