This study aimed to create and validate metagenomic libraries for sequencing to establish baseline understanding of active layer and permafrost microbial community functional gene composition and observe the changes that occur in functional gene abundance in these microbial communities when permafrost soils thaw. The soils used in this study were obtained from Cambridge Bay, Nunavut in the Canadian High Arctic. Five different thaw treatments, differing in soil compositions of permafrost and active layer soil were prepared in triplicate and incubated at 8 °C for 12 weeks. These treatments consist of permafrost and active layer soil incubated on their own and three other treatments with varying mixtures of both soils, designed to test how they will respond to permafrost thaw. Microbial communities were expected to change uniquely in their respective treatments over the experimental period. DNA and RNA extractions were performed on pre-thaw samples and samples after 12 weeks of incubation. Obtained DNA concentrations ranged from 93.5-425ng/2g of soil and extracted RNA samples were too low to pursue sequencing. DNA libraries were successfully prepared for Illumina sequencing (as assessed by size distribution analysis, confirmed by gel electrophoresis and Bioanalyzer assays), library quantification using a Qubit fluorometric assay, and qPCR analysis. Size distribution ranged from 310-570bp, and library concentrations were found to range from 0.020-53.46nM. Metagenomic sequencing of these libraries will allow insight into the functional gene abundance and microbial community changes that occur as these soils thaw.
