4.2.2. Gene Expression Analysis

Total RNA was extracted from placental tissues (*n* = 20 control and *n* = 20 PE) using RNeasy Kits (Qiagen, Glen Forrest, Australia) and reverse transcribed to form cDNA with SuperScript III Reverse Transcriptase (Invitrogen, ThermoFisher Scientific, Scoresby, Australia), according to manufacturer's protocol. mRNA expression of the creatine synthesizing enzymes (*GATM*-the gene that expresses AGAT & *GAMT*), the creatine transporter (*SLC6A8*) and creatine kinase isoforms (*BBCK* & *CKMT1A*) was determined using Fluidigm Biomark HD system with TaqMan chemistry. TaqMan probe sequences are detailed in Table 2. *RN18S* was used as the housekeeping gene, after conserved expression between the cohorts was validated. Data from qPCR was analyzed according to the ΔΔ *C*T method [48] and results are expressed relative to the control cohort.


**Table 2.** TaqMan probe Sequences.
