Insight into soil microbial ecology through the development and application of serial analysis of ribosomal sequence tags (SARST) and a community-specific microarray ...

• Main Author: Neufeld, Josh David
• Format: Text
• Language: English
• Published: University of British Columbia 2009
• Subjects: Arctic; Tundra
• Online Access: https://dx.doi.org/10.14288/1.0092359; https://doi.library.ubc.ca/10.14288/1.0092359

In order to overcome technical hurdles hindering progress in soil microbial ecology, this project involved the development and application of a molecular assay for measuring microbial community composition and diversity. Serial analysis of ribosomal sequence tags (SARST) generates libraries of a short and variable portion of the 16S rRNA gene. A series of enzymatic reactions amplifies and ligates ribosomal sequence tags (RSTs) into concatemers that are cloned and sequenced. On average, 5-10 RSTs were obtained from multiple phylotypes with each sequencing reaction. SARST was initially tested on: 1) activated sludge, 2) boreal forest soil, 3) a mixture of pure cultures and 4) duplicate libraries from an arctic tundra soil sample. SARST was also used to study microbial communities within soils sampled from Canada and Spain. Bacterial diversity of geographically distinct locations was characterized for Canadian arctic tundra and boreal forest soil biomes. Composite samples taken from arctic tundra demonstrated ...