2.4 RNA extraction
Total RNA was extracted from the entire CNS, and both eyes combined for
each squid (ncurrent-day CO2 = 10,
nelevated CO2 = 10 for each tissue) (Figure 1). Each
tissue sample was homogenised in RLT-Plus Buffer (Qiagen) with sterile
zirconia/silica beads (1mm diameter, BioSpec Products) in a
Mini-BeadBeater 96 (BioSpec Products) for a total of 2 minutes. Total
RNA was extracted using an AllPrep DNA/RNA Mini Kit (Qiagen). RNA
integrity of all 40 samples was measured on an Agilent 2200 TapeStation
(High Sensitivity RNA ScreenTape, Agilent) (Supplementary File S1).