Total RNA was extracted from individual tissues (3 female gonads, 3 male gonads, 4 gills, 4 digestive glands, 4 posterior adductor muscles, 4 labial palps, 4 mantles) or from pools of 6 individuals (4 pools of visceral ganglions and 3 pools of hemocytes) with Tri-Reagent kit (Sigma) and subsequently cleaned with Nucleospin RNA Clean-Up (Macherey Nagel) isolation columns.
The animals were dissected and the cephalic portion was removed, and the crania were subsequently cleaned with a 3% KHO solution [26].
Recovery time is the time required by a sensor so as to return to 90%% of the original baseline signal when the target gas is removed and the sensor is subsequently cleaned with dry air [29, 30, 31, 32, 33, 34].
The fluorine-doped tin oxide (FTO, <14 Ω/sq, 2.2-mm thick, Pilkington, Solar Energy Technology Co , Ltd ,uhan Jinge, China) substrates are first etched WuhanZn powder (Sigma Aldrich, St. Louis, MO, USA) and hydrochloric (HCl) acid (4 M, Sigma Aldrich) to form the desired pattern, which are subsequently cleaned with soap and ethanol (Sigma Aldrich).
One sample was prepared without special pretreatment, which means the surfaces were only rinsed with acetone and subsequently cleaned in an ultrasonic bath with isopropanol.
Sheared DNA fragments were subsequently cleaned with the DNA Clean & Concentrator-25 Kit (Zymo Research) and a fragment size distribution ranging from 300 bp to 500 bp was verified via Bioanalyzer (Agilent).
Samples were subsequently cleaned with a cation exchange Cartridge Kit (Applied Biosystems).
RNA was extracted from triplicate cultures using the hot-phenol method and subsequently cleaned up further on columns with the QiaGen RNA purification Kit.
Total RNA was treated with DNAse I (Promega Madison, WI) and subsequently cleaned using the RNeasy Mini Kit (Qiagen, Valencia, CA) as described by the manufacturer.
