Bee Nucleic Acid Extraction Protocol

Each sample was processed as a pool. Ten bees were placed in a 2mL microtube with 300 µL of DNA/RNA Shield (Zymo Research, Irvine, CA, USA) and crushed with a TissueLyser II (Qiagen, Hilden, Germany) for 3 min at 30 Hz, as previously reported [60 (link),61 (link)]. The obtained suspension was split into two aliquots, from which DNA and RNA were separately extracted. The extraction of the nucleic acids was performed with the Quick DNA Microprep Plus Kit (Zymo Research) and Quick RNA Microprep Plus Kit (Zymo Research). During the process, the modified manufacturer’s instructions for solid tissue processing were followed [19 (link),62 (link)]. The obtained nucleic acids were eluted in 100 µL of DNAase-RNase-free water and the extracts were stored at −80 °C until the qPCR assays.