2.5 Compound eyes sampling, RNA, and protein extraction
After the exposure experiment, the fresh compound eye mixture of 3O. oratoria s in each tank were used for mRNA extraction, and the
compound eye mixture of the remaining 3 O. oratoria s were used
for total protein extraction. The mRNA extraction processes were as
follows: Liquid nitrogen is added to the individual sample and fully
ground, and the high-quality total RNA (The RNA integrity number
detected by Agilent 2100 bioanalyzer is greater than 7) was extracted
using the standard Trizol Reagent Kit (Sangon Biotech). Subsequently, we
use RNA purification Beads (Illumina) to adsorb the mRNA from total RNA
and then use Beads Binding Buffer (Illumina) to clean the adsorbed mRNA
three times. The eluted mRNA was incubated at 94°C for 8 minutes to
achieve mRNA purification. The total protein extraction processes were
as follows: SDS lysate (Macklin) was added to the individual sample that
were fully ground with liquid nitrogen to promote complete sample
cleavage. PMSF (ThermoFisher) was added to the lysed sample to inhibit
protein hydrolysis. The solution was ultrasonically broken 3 times on
ice, and the total protein was then obtained in the supernatant by
centrifugation.