4.3. Sequencing of Microbiota in Cecal Pellets

DNA was extracted from mouse cecal pellets preserved in DNA/RNA Shield (Zymo Research, Irvine, CA, USA) utilizing Qiagen’s PowerFecal Pro DNA kit (Qiagen, Hilden, Germany) following manufacturer’s protocol. Extracted DNA was quantified with the ONE dsDNA QuantiFluor Dye System on Quantus Fluorometer (Promega, Madison, WI, USA). Following quantitation, purified sample DNA was normalized for downstream tagmentation and library fabrication utilizing Nextera DNA Flex kit (Illumina, San Diego, CA, USA). Prepared and indexed libraries were then pooled for next generation sequencing (NSG) with NextSeq 500/550 Mid-Output Kit v2.5 (300 cycles) (Illumina).