An improved method for intracellular DNA (iDNA) recovery from terrestrial environments

The simultaneous extraction of intracellular DNA (iDNA) and extracellular DNA (eDNA) can be improved by using a highly concentrated sodium phosphate buffer with a detergent mix  or EDTA. This improved protocol allows for higher recovery of iDNA from various environmental samples, including low-biomass iron-bearing rock samples, while buffers based solely on sodium phosphate are recommended for studies focusing on eDNA. These modifications may contribute to a more accurate characterization of modern and past ecosystems in environmental studies AbstractThe simultaneous extraction of intracellular DNA (iDNA) and extracellular DNA (eDNA) can help to separate the living in situ community (represented by iDNA) from background DNA that originated both from past communities and from allochthonous sources. As iDNA and eDNA extraction protocols require separating cells from the sample matrix, their DNA yields are generally lower than direct methods that lyse the cells within the sample matrix. We, therefore, tested different buffers with and without adding a detergent mix (DM) in the extraction protocol to improve the recovery of iDNA from surface and subsurface samples that covered a variety of terrestrial environments. The combination of a highly concentrated sodium phosphate buffer plus DM significantly improved iDNA recovery for almost all tested samples. Additionally, the combination of sodium phosphate and EDTA improved iDNA recovery in most of the samples and even allowed the s...
Source: MicrobiologyOpen - Category: Microbiology Authors: Tags: ORIGINAL ARTICLE Source Type: research