Seq of honeybee hemolymph
Honeybee hemolymph samples underwent filtration using a 0.8 m polyethersulphone filter (Vivaclear, Sartorius) and centrifugation at 2,000 g for 5 minutes. To ensure quality control and semi-quantification in downstream data analysis, a spike-in virus was introduced. The resulting filtrates underwent enzymatic host depletion and ad random amplification. The obtained DNA was then subjected to rapid library preparation using the RBK096 library prep (ONT), allowing multiplexing of up to 24 samples per run. Sequencing was performed on R9.4.1 flow cells (ONT) on a GridION device. This facilitated real-time data acquisition, super accurate base calling, and demultiplexing through Guppy (v.6.1.5; ONT).