Single-cell whole genome sequencing of bacteria from saliva
Single-cell whole genome sequencing of bacteria from saliva
Fresh saliva was obtained from a healthy (non-patient) human donor (ethical permit # 08-047M). The saliva (5mL) was collected in a 50mL falcon tube and vortexed for 3 min with glass beads (Fisherbrand, catalog #41401001). The sample was subsequently filtered using a 35μm filter (Cat no. 352235, Corning). The filtrate was further centrifuged at 5,000g for 10 minutes. The supernatant was discarded and the pellet was washed with 1mL of PBS three times. Finally, the pellet was resuspended in 10% glycerol and stored at -80°C. Nucleic acid was extracted according to lysis protocols R1, R2, R3 described further in our manuscript Libraries were prepared using SPCs as specified in our manuscript