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Evaluation and optimization of nucleic acid extraction methods for the molecular analysis of bacterial communities associated with corroded carbon steel
Marty, F.; Ghiglione, J.-F.; Païssé, S.; Gueuné, H.; Quillet, L.; van Loosdrecht, M.C.M.; Muyzer, G. (2012). Evaluation and optimization of nucleic acid extraction methods for the molecular analysis of bacterial communities associated with corroded carbon steel. Biofouling (Print) 28(4): 363-380. http://dx.doi.org/10.1080/08927014.2012.672644
In: Biofouling. Taylor & Francis: Chur; New York. ISSN 0892-7014; e-ISSN 1029-2454, meer
Peer reviewed article  
Beschikbaar in  Auteurs 
Trefwoorden
    Acids > Organic compounds > Organic acids > Nucleic acids
    Carbon steels
    Marien/Kust
Author keywords
    Nucleic acid extraction; Microbiologically influenced corrosion; Molecular fingerprinting,
Auteurs  Top 
  • Marty, F.
  • Ghiglione, J.-F.
  • Païssé, S.
  • Gueuné, H.
  • Quillet, L.
  • van Loosdrecht, M.C.M.
  • Muyzer, G., meer
Abstract
    Different DNA and RNA extraction approaches were evaluated and protocols optimized on in situ corrosion products from carbon steel in marine environments. Protocols adapted from the PowerSoil DNA/RNA Isolation methods resulted in the best nucleic acid (NA) extraction performances (ie combining high NA yield, quality, purity, representativeness of microbial community and processing time efficiency). The PowerSoil RNA Isolation Kit was the only method which resulted in amplifiable RNA of good quality (ie intact 16S/23S rRNA). Sample homogenization and hot chemical (SDS) cell lysis combined with mechanical (bead-beating) lysis in presence of a DNA competitor (skim milk) contributed to improving substantially (around 23 times) the DNA yield of the PowerSoil DNA Isolation Kit. Apart from presenting NA extraction strategies for optimizing extraction parameters with corrosion samples from carbon steel, this study proposes DNA and RNA extraction procedures suited for comparative molecular analysis of total and active fractions of bacterial communities associated with carbon steel corrosion events, thereby contributing to improved MIC diagnosis and control.
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