Amplicon sequencing of Porcilis MLV-DV
For whole genome amplification, a tailed tiling amplicon scheme was designed, and 34 amplicons that theoretically covered the full-length Porcilis MLV-DV genome were generated. Whole genome amplification connected with library preparation consisted of a two-step PCR protocol after the cDNA synthesis. Random based reverse transcription was performed using the SuperScript IV VILO Master Mix according to the manufacturers instruction. Both PCR rounds (PCR-1 and PCR-2) were performed using Q5U Hot Start High-Fidelity DNA Polymerase according to the manufacturers instruction. PCR-1 was performed to amplify the genome of Porcilis MLV-DV, and in this step the Illumina-compatible sequencing primer binding sites were added to the PCR products. During the second PCR, Illumina specific adapters and indices were added to the PCR-1 products. The amplicon libraries were sequenced on Miseq using the Reagent Kit v3 (600-cycle).