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

• DOI: 10.1002/mbo3.1369
• Publication Date: 2023-06-26T05:41:08Z
• Authors: Diego Medina Caro; Lucas Horstmann; Lars Ganzert; Romulo Oses; Thomas Friedl; Dirk Wagner
• Keywords: 500; Original Articles; DNA; cell detachment; DNA desorption; Microbiology; QR1-502; Phosphates; deep biosphere; intracellular DNA; low biomass; DNA extraction; Ecosystem; Edetic Acid

The simultaneous extraction of intracellular DNA (iDNA) and extracellular (eDNA) can help to separate the living in situ community (represented by iDNA) from background that originated both past communities allochthonous sources. As iDNA eDNA protocols require separating cells sample matrix, their yields are generally lower than direct methods lyse within matrix. We, therefore, tested different buffers with without adding a detergent mix (DM) protocol improve recovery surface subsurface samples covered variety terrestrial environments. combination highly concentrated sodium phosphate buffer plus DM significantly improved for almost all samples. Additionally, EDTA most even allowed successful extremely low-biomass iron-bearing rock taken deep biosphere. Based on our results, we recommend using either (NaP 300 mM + DM) or mM). Furthermore, studies rely pool, solely based because addition resulted decrease These improvements reduce bias environmental contribute better characterizations modern ecosystems.