Quantification of KSHV Genome Copy Number

Quantification of KSHV genome copy number was carried out as previously described [54 (link)]. In brief, BCBL1-Tet-K-RTA were treated with Dox (1 μg/ml) and sodium butyrate (0.5 mM) for 48 h to induce lytic reactivation, and the supernatant (500 μl) was collected and treated with 7.5 U of DNase I (Solarbio, Beijing, China) for 1 h at 37°C. Then 30 μl of proteinase K (20 mg/ml, Solarbio, Beijing, China) and 50 μl of 20% SDS was added into the mixture. After incubation at 65°C for 1 h, genomic DNA was extracted by phenol-chloroform extraction and the DNA pellet was resolved in 50 μl of TE buffer. The genomic DNA was diluted 20 times and KSHV genomic DNA was quantified by qPCR. A stand curve was generated using serial dilutions of a pEF-FLAG-K-RTA plasmid. The primers used for the quantification are provided in S2 Table.