Urinary Microbiota Associated with Preterm Birth
Maternal urineĘ(60mL) specimens were collected into 120cc plastic containers, aliquoted, and kept cold (2-40C) and frozen (-700C) within 1 hour of collection.  Participants were provided with Benzalkonium (BZK) wipes and instructed to wipe from front to back and then collect the urine.  2mL aliquots were mailed on dry ice to the laboratory of Piyathilake and kept frozen at -700C until microbial DNA was isolated.Microbial DNA was isolated using the Fecal DNA isolation kit from Zymo Research (Irvine, CA). The V4 region of the 16S rRNA gene (515F/806R) was amplified from extracted DNA using the protocol and region-specific PCR primers. PCR products were purified by gel electrophoresis and quantitated using the Picogreen (Invitrogen) dsDNA quantitation assay prior to sequencing. Paired end sequencing (2 x 250) was performed on the Illumina MiSeq platform.