DNA Extraction for Streamlined Metagenomics of Diverse Environmental Samples
Author(s) -
Clarisse Marotz,
Am Amir,
Greg Humphrey,
James Gaffney,
Grant Gogul,
Rob Knight
Publication year - 2017
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000114559
Subject(s) - metagenomics , dna extraction , replicate , amplicon , biology , environmental dna , extraction (chemistry) , amplicon sequencing , computational biology , polymerase chain reaction , 16s ribosomal rna , chromatography , genetics , gene , ecology , biodiversity , chemistry , mathematics , statistics
A major bottleneck for metagenomic sequencing is rapid and efficient DNA extraction. Here, we compare the extraction efficiencies of three magnetic bead-based platforms (KingFisher, epMotion, and Tecan) to a standardized column-based extraction platform across a variety of sample types, including feces, oral, skin, soil, and water. Replicate sample plates were extracted and prepared for 16S rRNA gene amplicon sequencing in parallel to assess extraction bias and DNA quality. The data demonstrate that any effect of extraction method on sequencing results was small compared with the variability across samples; however, the KingFisher platform produced the largest number of high-quality reads in the shortest amount of time. Based on these results, we have identified an extraction pipeline that dramatically reduces sample processing time without sacrificing bacterial taxonomic or abundance information.
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