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Evaluation of Lysis and Recovery of Genomic DNA from Microbiota to Maximize Efficiency for DNA Sequencing
Author(s) -
Proctor Caleb McKinley,
Okparanta Akelachi,
Ryan Gabriella,
Easparro Brandon,
Nash Rodney
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.06002
Subject(s) - biology , microorganism , lysis , microbiome , dna sequencing , soil microbiology , population , microbial ecology , computational biology , microbiology and biotechnology , ecology , dna , genetics , bacteria , soil water , demography , sociology
Microbes, specifically microbes found in mixed populations in guts, soil, feces, and other tissues, have been the object of much research for commensal relationships for decades. Many of these microbes are ubiquitous across many species or biomes but are present very in low quantities due to the inherent challenges of microorganism survival in environmental conditions. While, in some cases, only present in low quantity, soil microorganisms are a critical component of our ecosystem. Soil microbiome studies aim to characterize the microbial repertoire and to correlate the microbial population with soil and ecological functions. Given the time involved and complexity of culture‐based enrichments, along with the inability to grow some species in culture, many modern studies involve direct cell lysis and DNA recovery from soil followed by PCR or genetic sequencing for target enrichment, detection, and analysis. In order to evaluate the organisms, present in such low quantities, methods must be developed to lyse all organisms present in soil of various sizes and toughness’. In this study we evaluate the lysis and recovery of a microbiological standard using commercially available kits and genetic sequencing to show an ideal bead milling matrix for microbiota in soil. Support or Funding Information Funding for this project was provided by Omni International