Koala chlamydia detection protocol

Genomic DNA (DNA) was extracted from koala urogenital and ocular swabs using QIAamp DNA minikit (Qiagen, Valencia, CA) as previously described^{41 (link)}. The extracted DNA was screened for the presence of C. pecorum using a species-specific quantitative real-time PCR (qPCR) that targeted the 16 S rRNA gene^{41 (link)} (Supplementary Table S6). The reaction mix contained: 5–50 ng DNA; 1x QuantiTect SYBR Green PCR Master Mix (Qiagen, Valencia, CA); and 10 µM of each primer made up to a final volume of 25 µL with molecular grade water. Thermocycling was performed on a RotorGeneQ 5-plex HRM platform (Qiagen, Valencia, CA) using a 15-minute initial denaturation at 94 °C followed by 40 cycles at: 94 °C for 15 seconds; 57 °C for 30 seconds; 72 °C for 25 seconds for amplification.