16S rRNA sequencing of urinary tract microbiome in patients undergoing dialysis and patients after kidney transplantation
A urine sample was obtained by a sterile mid-stream collection. Derived sample was centrifuged at 800 rcf for 10 minutes and the pellet was stored at -80C. Genomic microbial DNA was extracted from urine pellet using Genomic Mini AX Body Fluids extraction kit (A&A Biotechnology), according to the manufacturer's protocol with one modification, in a form of adding 10 l of each of mutanolysin, lysostaphin, lysozyme, and lyticase to ensure cell lysis. V3 and V4 hypervariable regions of the bacterial 16S rRNA gene were amplified using 515F and 806R primers. Libraries were prepared with Nextera XT Index Kit followed by sequencing using MiSeq System.