16S rDNA sequencing, Studies of Effects of Fecal Microbiota Transplantation on ADHD-like model rat.

In this project, we used 16S rDNA analysis to demonstrate that attention deficit hyperactivity disorder (ADHD)-like model rats have a gut microbiota composition distinct from that of healthy rats. Furthermore, we showed that fecal microbiota transplantation (FMT) from healthy rats improved hyperactivity and altered the gut microbiota in ADHD-like model rats. We submitted the 16S rDNA sequences of 6-week-old healthy and ADHD-like model rats, as well as 8-week-old ADHD-like model rats that underwent FMT at 6 weeks of age from healthy rats. The sequences were determined as follows: Genomic DNA was extracted from fecal samples using the NucleoSpin Tissue kit (Macherey-Nagel, Germany), and the quality was assessed with a Qubit Flex Fluorometer (Invitrogen, Carlsbad, CA, USA). The 16S rRNA gene in the V3-V4 region was amplified using the primers 341F and 805R with 2X KAPA HiFi HS ReadyMix (Kapa Biosystems, Wilmington, USA). Following purification and quantification of the PCR amplicons using Sera-Mag Select magnetic beads (Cytiva, Tokyo, Japan), sequencing libraries were prepared using the Nextera XT Index Kit v2 Set A (Illumina, San Diego, CA, USA) and sequenced on the Illumina MiSeq platform.