Fabrication of DNA Microarrays by Digital Solid-Phase PCR in a Next Generation Sequencing Chip

Jochen Hoffmann

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Doctoral Thesis / Dissertation from the year 2013 in the subject Biology - Genetics / Gene Technology, grade: 1,0, University of ...

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Doctoral Thesis / Dissertation from the year 2013 in the subject Biology - Genetics / Gene Technology, grade: 1,0, University of Freiburg (Institut für Mikrosystemtechnik), course: Mikrosystemtechnik, language: English, abstract: A process for the fabrication of DNA microarrays by copying a next-generation sequencing chip is presented in this thesis. DNA molecules of a DNA library are mixed with a biochemical reaction mix and loaded into hundreds of thousands wells of the commercial sequencing chip PicoTiterPlate (PTP, Roche). The DNA randomly distributes onto the wells so that statistically one well contains one single DNA molecule. All wells of the PTP are closed with a microscope slide and the DNA is amplified by polymerase chain reaction (PCR). Each single PCR product is attached as a DNA cluster in the PTP and on the corresponding position on the slide since both surfaces have been coated with PCR primers. The sequence of each DNA cluster can be obtained by a subsequent sequencing reaction in the PTP. This also reveals the sequences of each DNA cluster on the slide thus allowing the slide to be used as a standard DNA microarray.