Healthy volunteer samples were collected and total RNA purified using the RNeasy PowerMicrobiome Kit. Profiling and quantification of 1 µl of RNA was performed using an Agilent Bioanalyzer RNA Nano Kit chip. In the electropherogram, clear 16S and 23S bands are shown for prokaryotic rRNA. RNA processed through the RNeasy PowerMicrobiome Kit was demonstrated to be high-quality by an RIN of 9.4. (Data kindly provided by C. Vincent, McGill University, Montreal, Canada.)
Experimental RNA preservation solution was used to preserve fresh stool samples at room temperature or on ice. Collection and processing occurred within the same 24-hour period. RNeasy PowerMicrobiome Kit was used to isolate total RNA. Each sample (10 µl per) was examined on a 1% agarose gel.