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Exploring the fusion of metagenomic library and DNA microarray technologies

Posted on:2007-07-18Degree:M.ScType:Thesis
University:McGill University (Canada)Candidate:Spiegelman, DanFull Text:PDF
GTID:2450390005487497Subject:Biology
Abstract/Summary:
We explored the combination of metagenomic library and DNA microarray technologies into a single platform as a novel way to rapidly screen metagenomic libraries for genetic targets. In the "metagenomic microarray" system, metagenomic library clone DNA is printed on a microarray surface, and clones of interest are detected by hybridization to single-gene probes. This study represents the initial steps in the development of this technology. We constructed two 5,000-clone large-insert metagenomic libraries from two diesel-contaminated Arctic soil samples. We developed and optimized an automated fosmid purification protocol to rapidly-extract clone DNA in a high-throughput 96-well format. We then created a series of small prototype arrays to optimize various parameters of microarray printing and hybridization, to identify and resolve technical challenges, and to provide proof-of-principle of this novel application. Our results suggest that this method shows promise, but more experimentation must be done to establish the feasibility of this approach.
Keywords/Search Tags:DNA, Metagenomic library, Microarray
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