*2.3. RNA Isolation and cDNA Synthesis*

Tissues were taken out of RNAlater and cleaned with sterile pipettes. *N. furzeri* (NFu) total RNA was isolated from 10 animals with QIAzol (Qiagen), according to a modified manufacturer's protocol [27]. Homogenization was performed using a TissueLyzer II (Qiagen) at 20 Hz for 2–3 × 1 min. Total RNA was then quantized with Eppendorf BioPhotometer. Then, 500 ng of each sample were retrotranscribed to cDNA in a 20 μL volume, using the QuantiTect® Reverse Transcription Kit (Qiagen), following the supplier's protocol. Newly synthetized cDNAs were then diluted to a final volume of 200 μL with ultra-pure sterile water to an approximate final cDNA's concentration of 40 ng/μL.
