Exploring the fusion of metagenomic library and DNA microarray technologies

• Main Author: Spiegelman, Dan.
• Format: Thesis
• Language: English
• Published: McGill University 2006
• Subjects: Microbial genomes; Gene libraries; DNA microarrays; Soil microbiology > Nunavut > Ellesmere Island; Oil pollution of soils > Nunavut > Ellesmere Island; Arctic; Ellesmere Island; Nunavut
• Online Access: http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=98805

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.