title: Modified low-salt CTAB extraction of high-quality DNA from contaminant-rich tissues.
abstract: The increasing use of high-throughput sequencing platforms has made the isolation of pure, high molecular weight DNA a primary concern for studies of a diverse range of organisms. Purification of DNA remains a significant challenge in many tissue and sample types due to various organic and inorganic molecules that coprecipitate with nucleic acids. Molluscs, for example, contain high concentrations of polysaccharides which often coprecipitate with DNA and can inhibit downstream enzymatic reactions. We modified a low-salt CTAB (MoLSC) extraction protocol to accommodate contaminant-rich animal tissues and compared this method to a standard CTAB extraction protocol and two commercially available animal tissue DNA extraction kits using oyster adductor muscle. Comparisons of purity and molecular integrity showed that our in-house protocol yielded genomic DNA generally free of contaminants and shearing, whereas the traditional CTAB method and some of the commercial kits yielded DNA unsuitable for some applications of massively parallel sequencing. Our open-source MoLSC protocol provides a cost-effective, scalable, alternative DNA extraction method that can be easily optimized and adapted for sequencing applications in other contaminant-rich samples.
authors: Department of Fisheries and Oceans Canada, Gulf Fisheries Center, 343 Universite Avenue, Moncton, NB, E1C 9B6, Canada.
authors: Jean-René
authors: JR
authors: Arseneau
authors: Department of Fisheries and Oceans Canada, Gulf Fisheries Center, 343 Universite Avenue, Moncton, NB, E1C 9B6, Canada.
authors: Royce
authors: R
authors: Steeves
authors: Department of Fisheries and Oceans Canada, Gulf Fisheries Center, 343 Universite Avenue, Moncton, NB, E1C 9B6, Canada.
authors: Mark
authors: M
authors: Laflamme
mesh_terms: D000818:Animals; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry*; D004247:DNA / Q000302:isolation & purification*; D010125:Ostreidae; D012492:Salts
chemical_list: D002593:Cetrimonium Compounds; D012492:Salts; D004247:DNA; D000077286:Cetrimonium
keywords: DNA extraction; genomics; high-throughput sequencing; invertebrates; molluscs; polysaccharide