*2.5. Genotyping*

We collected saliva samples from each pregnan<sup>t</sup> woman participating in the study, and genomic DNA was isolated from buccal epithelial cells using the Aidar and Line (2007) protocol [32]. FTO (rs9939609 and rs17817449) and ADRB2 (rs1042713 and rs1042714) polymorphisms were genotyped by real-time PCR using TaqMan® assays (Thermo Fisher Scientific, Carlsbad, CA, USA). Reactions were performed in 10-μL volumes containing DNA (2 μL), Universal Master Mix (5 μL), TaqMan Genotyping Assay specific for each polymorphism (0.25 μL), and MiliQ (2.75 μL). Amplification was carried out in a StepOne® Plus Real-Time PCR System (ThermoFisher) according to the manufacturer's recommendations for the number of cycles and temperatures. Negative and positive controls were included in the plate.
