ASF Virus Pol19_53050_C1959/19 Isolation

The ASF virus Pol19_53050_C1959/19 was obtained on the 5th day of the 3rd passage in PPAM cell culture. Then, 2 × 200 µL of the cell culture was subjected to DNA extraction with a QiAMP DNA Mini Kit (Qiagen, Hilden, Germany). Final elution of DNA from the column was performed in triplicate (3 × 30 µL). Obtained DNA (2 × 90 µL) was pooled and cleaned-up using KAPA Hyper Pure Beads (Roche, Basel, Switzerland), and recovered in 20 µL of DNAse/RNAse free water. The final concentration of total DNA was measured with Nanodrop (Thermo Fisher Scientific, Waltham, MA, USA) (15.5 ng/µL), A260/A280 factor reached 1.86. The dsDNA concentration was measured using a Qbit dsDNA High Sensitivity kit (Thermo Fisher Scientific, Waltham, MA, USA) and reached 8.52 ng/µL. The remaining three isolates were prepared as described earlier [2 (link)].