mNGS of Homo sapiens: Plasma
Five-ten mL of whole blood was collected from each patient and stored in an EDTA tube at 4 C. Samples were centrifuged within 1h after collection for 10 min at 1600 x g at 4 C. Plasma samples were transferred to 1.5-mL microcentrifuge tubes and stored at -80C until DNA extraction. Nucleic acids were extracted from 0.3 mL plasma, and DNA libraries were established by shearing, end repair, A-tailing, adapter ligation, and PCR amplification. DNA libraries were denatured and circularized. DNA nanoballs were generated with the ssDNA circle by rolling circle replication. Finally, sequencing was performed on a BGI MGISEQ-2000 platform in the SE50 format.