Evaluation of five in situ lysis protocols for PCR amenable metagenomic DNA from mangrove soils

• Main Authors: Harisree P. Nair, Helvin Vincent, Sarita G. Bhat
• Format: Article
• Language: English
• Published: Elsevier 2014-12-01
• Series: Biotechnology Reports
• Subjects: Metagenome; In situ lysis; DNA extraction
• Online Access: http://www.sciencedirect.com/science/article/pii/S2215017X14000423

Microbes in nature are rarely amenable to growth by standard microbiological methods, with the majority being unculturable. Metagenomic methods help to bypass and overcome the limitations of traditional culturing method; wherein total community DNA is isolated, cloned into suitable vector and host systems. However, isolation of total community DNA itself remains a challenge. In this study five methods of total community DNA isolation from three different mangrove soils were evaluated to test its PCR amenability. The yield and purity of the isolated DNA was also analysed. The quantity of DNA by all 5 methods although reasonably high, contained residual humic contaminants. Of the five, the method employing liquid nitrogen yielded readily amplifiable DNA, while that by all others required further downstream processing to achieve purity and PCR amenability.