Methylation procedures affect PLFA results more than selected extraction parameters

Microorganisms are key players in organic matter and nutrient cycles of terrestrial ecosystems. The analysis microbial membrane lipids, phospholipid fatty acids (PLFAs) has strongly improved our understanding how processes contribute to these cycles. proven yield robust results, but adaptations analytical parameters laboratory needs might lead pitfalls impede comparability PLFA results between different studies. Here, we show a set four (freeze-drying vs. field moist, amount sample extracted, age solvent mixture, methylation methods) influence the quantitative qualitative analysis. Freeze-drying moist samples extracted had only minor effects on concentrations recovery community structure. Nevertheless, important consider, especially if treatment an experiment expected be low. use weeks old extraction solution resulted 12% lower as well significant differences relative abundance functional groups. This suggests that should prepared day or components added sequentially sample. Most importantly, choice method led both, (35%) groups, making comparisons studies difficult. Our study provides valuable ranking need considered during implementation also highlights fact using methods limited.