Cfx pcr instruments

Fetal membranes were dissociated in Trizol (Life Technologies, Carlsbad, CA, USA) using a high-speed homogenizer. RNA was isolated using an RNeasy Kit (QIAGEN, Hilden, Germany) and treated with RNase-free DNase (QIAGEN). cDNA was synthesized using a High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher, Waltman, MA, USA) according to the manufacturer’s instructions. Real-time qRT-PCR was performed using validated TaqMan gene expression NHERF1 primer (assay ID: Hs00188594_m1, gene symbol: SLC9A3R1), IL-10 primer (assay ID: Hs00961622_m1, gene symbol: IL-10), IL-6 primer (assay ID: Hs00174131_m1, gene symbol: IL-6), and TNF-α (assay ID: Hs00174128_m1, gene symbol: TNF) and probe sets according to the manufacturer’s instructions. qRT-PCR reactions were carried out using Bio-Rad CFX PCR Instruments (Bio-Rad, Hercules, CA, USA). Results were analyzed using CFX maestro software. mRNA expression was calculated using the 2^−ΔCt method with GAPDH as an internal reference control.

A high-speed homogenizer dissociated FMs and placentas in Trizol (Life Technologies, Carlsbad, CA, USA). RNA was isolated using the RNeasy Kit (QIAGEN, Hilden, Germany) and treated with RNase-free DNase (QIAGEN). According to the manufacturer’s instructions, cDNA was synthesized using a High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher, Waltman, MA, USA). According to the manufacturer’s instructions, real-time qRT-PCR was performed using a validated TaqMan gene expression ABCG2 primer (assay ID: Hs00188594_m1, gene symbol: ABCG2) probe sets. qRT-PCR reactions were carried out using Bio-Rad CFX PCR instruments (Bio-Rad, Hercules, CA, USA). Results were analyzed using CFX Maestro software. The gene expression was calculated using the 2−∆Ct method with GAPDH as an internal reference control.