Qiashredder protocol

HSV-1 (GFP strain) was propagated in VERO cells as described (38 (link)). OKF-6/TERT-1 cells were infected at an MOI of 0.1:1 for 24 h. After infection, cells were harvested and homogenized using Qiagen QIAshredder protocol. The obtained lysate was then used to extract RNA and DNA using the Qiagen RNeasy Plus Micro kit. Once RNA was extracted using the manufacturer protocol, 50 µl of 8.0 mm NaOH was added directly to membrane. Tubes were then incubated for 10 min at 55°C. DNA was eluted by centrifugation for 3 min at 5,000 × g. qPCR was performed on a BIO RAD myCycler using SsoAdvanced Universal SYBR green supermix (Bio-RAD #1725274), using 1.5 µl cellular DNA per reaction, with HSV UL30 primers (F: 3′-AGAGGGACATCCAGGACTTTGT-5′; R: 3′-CAGGCGCTTGTTGGTGTAC5′). DNA levels were normalized to β-actin in each sample and the levels determined using the 2^−ΔΔCT method compared to ethanol-treated cells.

Cells were harvested and homogenized using Qiagen QIAshredder protocol. The obtained lysate was then used to extract RNA and DNA using the Qiagen RNeasy Plus Micro kit. Once RNA was extracted using the manufacturer protocol, DNA was extracted as described [49 (link)]. Using the same RNeasy spin columns 50 µL of 8.0 mM NaOH was added directly to membrane. The tubes were then incubated for 10 min at 55 °C. DNA was eluted by centrifugation for 3 min at 5000× g.