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Development and use of a microchip for molecular genotyping

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The DNA-polymeras catalyzed "mini-sequencing" single nucleotide primer extension reaction allows highly specific discrimination between genotypes. The comprehensive genotyping system involves (i) chemistry and printing robotics for making primer microarrays, (ii) multiplex PCR amplification of genomic DNA samples, (iii) optimally designed fluorescent mini-sequencing reactions on the arrays, and (iv) a system for measuring and interpreting the results. Microscope glass slides serve as the microarray. The mini-sequencing reactions are performed in 80 miniaturized reaction chambers in a 384- microtiter plate well format on each microscope slide. Using this system demonstration projects, in which altogether 65 different SNPs were analyzed producing 90,000 genotypes. The technology is promising for large scale and cost-effective genotyping of SNPs in disease genetics and pharmacogenetics. The result is a procedure for genetic analyses in a micorarray (chip) format. It belongs to the fields of biotechnology and molecular genetics. The field of micorarray technology for genetic analyses is a new one. The instruments required have only recently become commercially available. The result represents cutting edge technology. It is a combination of several steps that are utilized together in the procedure, but the steps may also be utilized separately.

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