Fig. 1

Workflow of 16S rRNA amplicon sequencing with the MinION platform and bioinformatic analysis. After collecting the vaginal sample, it should be stored in a freezer at − 30 °C until DNA extraction begins. Some swab collection kits should be stored at room temperature, in accordance with the manufacturer’s instructions. Sequencing libraries are generated by a four-primer PCR-based strategy; in the initial stages of PCR, the 16S rRNA gene is amplified with an inner primer pair. The PCR product is amplified with the outer primers and targeted to introduce identical barcode and tag sequences at both ends, allowing for the attachment of adapter molecules in a one-step reaction. The library is then loaded into a MinION connected to a personal computer. With our experimental method, sequencing runtime of 10 min is sufficient for 3000 reads to be obtained. A final report of the microbiota can be presented with a total analysis time of approximately 4 h