16S rRNA seq of human feces: HIV non-controler
Design: DNA extraction was performed with the commercial kit QIAamp DNA Stool Mini Kit (Qiagen, Hilden,Germany) and stored at -20 C. DNA was quantified with the Qubit dsDNA HS kit (Thermo Fisher Scientific, USA) and amplified with the Illumina primers for the V3 and V4 regions of the 16S rRNA. Amplification was confirmed through electrophoresis in agarose gel. For library preparation, PCR product was purified with AMPure XP (Beckman Coulter, Brea, USA), adaptors for sequence identification were added with the commercial kit Nextera XT Index (Illumina, San Diego, USA), and samples were quantified through qPCR with the KAPA Universal Library Quantification Kit (Illumina) commercial kit. After quantification, a 3 nM equimolar pool was prepared for sequencing.