Extraction and Quantification of Placental RNA

Total RNA was extracted both from placental tissue (approximately 100 mg obtained from PE and normotensive controls) and trophoblast cells after hypoxia/reperfusion using TRI reagent (Invitrogen, UK). Total RNA (1 μg) was reverse transcribed to cDNA using the GoScript™ Reverse Transcriptase System (Promega, USA) according to the manufacturer's instructions. Quantitative reverse transcription‐PCR (qRT‐PCR) was performed as previously described.
⁴³ (link) In brief, qRT‐PCR was carried out on a CFX qRT‐PCR System using a SYBR® Green PCR master mix detection kit (Promega, USA). Primer pairs are listed in Table 1. The relative gene expression was calculated using the 2^−ΔΔCq method, using tyrosine 3‐monooxygenase/tryptophan 5‐monooxygenase (YWHAZ) as the reference gene.