16S Stool male Control
DNA was obtained from 200 mg from stool samples from post-COV patients and controls by using QIAamp Fast DNA Stool Mini Kit (Qiagen, CA, USA), according to manufacturer instructions. For bacterial 16S sequencing, DNA was quantified by Quantus fluorometer and adjusted to 5 ng/mL using Tris buffer (10 mM, pH 8.5). V3 and V4 regions of the bacterial 16S were amplified by using bacterial DNA, V3/V4 primers and the 2X KAPA HiFi HotStart Ready Mix (Kapa Biosystems, MA, USA). PCR purification was performed using AMPure XP Beads Kit (BD Biosciences, CA, USA). DNA libraries were constructed according to the Illumina protocols and the sequencing was conducted by an Illumina MiSeq platform system.