An Optimization to Increase Bacterial DNA Yield in a Lunar Regolith Simulant

Abstract Microbiological methods to transform lunar regolith into a substrate for plant cultivation are important for a biological regenerative life support system (BLSS). This study presents a rapid, effective method for optimizing the isolation of bacterial DNA from a consortium inoculated into the lunar regolith simulant LHS-1. The protocol yielded up to 717 ± 221 ng of high-quality DNA, sufficient for 16S rRNA amplicon sequencing. The approach, based on bacterial suspension and extraction using PrepMan ™ Ultra reagent followed by Zymo® purification, overcomes low-yield issues typical in regolith-based samples. This method enables reliable taxonomic analysis, contributing to understanding microbial interactions in regolith environments relevant to space agriculture.