2.2 | Total RNA extraction
Total RNA was extracted using TriPure Isolation reagent (Roche, Base,
Switzerland) in conjunction with the PureLinkRNA Mini Kit (Thermo Fisher
Scientific, Carlsbad, CA, USA). In the laboratory, frozen zooplankton
samples homogenized with TriPure were slowly thawed on ice. During the
phase separation, chloroform added to each sample was adjusted
proportionally based on the TriPure used in the homogenization. For each
sample, 600 µl colourless upper aqueous phase was collected carefully
from the homogenate and used for the elution procedure. A total volume
of 200 µl of RNase-Free water was used for the final elution. The
integrity of the extracted RNA was assessed using a Bioanalyzer (Agilent
Technology, CA, USA). High integrity RIN (RNA Integrity Number) indexes
were observed for all the samples used in this study (Table S1), except
for one sample (sample ID: N3NW0 F1).