Viral metagenome of pigeons: PLK2
Total viral nucleic acid was subjected to extraction using a QIAamp MinElute Virus Spin Kit (Qiagen, Hilden, Germany), according to the manufacturers instructions. For the 24 pigeon faecal specimens, pools of extracted nucleic acids from eight faecal specimens were subsequently generated, resulting in three separate pools. The enriched viral nucleic acid preparations from the respective pools, non-specific RNA and DNA amplification, were then performed by random RT-PCR, using a primer with randomised octamer at the 3 ends. Finally, the amplified viral nucleic acid libraries were subjected to a library preparation step using a Nextera XT sample preparation kit, following the manufacturers instructions, and sequenced using a MiSeq platform with a MiSeq Reagent Kit version 2 (Illumina).