Optimization of RNA extraction for bacterial whole transcriptome studies of low-biomass samples
Tom Verbeelen, Rob Van Houdt, Natalie Leys, Ramon Ganigué, Felice Mastroleo
Abstract
were used as model organisms for optimization of the RNA isolation protocol. Enzymatic lysis through lysozyme digestion generated high-quality, high-yield RNA samples. Subsequent RNA-seq analysis resulted in qualitative data for both strains. The RNA extraction procedure is suitable for experiments with volume and/or biomass limitations, e.g., as encountered during space flight experiments. Furthermore, it will also result in higher RNA yields for whole transcriptome experiments where sample volume and/or biomass was increased to compensate the low-biomass characteristic of autotrophs.
Topics & Concepts
RNA extractionRNABiomass (ecology)LysisTranscriptomeExtraction (chemistry)Nitrosomonas europaeaNitrobacterChemistryFood scienceRibosomal RNABiologyChromatographyNitriteBiochemistryGene expressionGeneAgronomyEcologyNitrateGenomics and Phylogenetic StudiesMolecular Biology Techniques and ApplicationsMicrobial Community Ecology and Physiology