cDNA Synthesis and qRT-PCR Analysis

The cDNA synthesis for qPCR was transcribed through a OneScript cDNA Synthesis Kit (ABM, Richmond, ON, Canada). The levels of mRNA were quantified through quantitative real-time PCR (qRT-PCR) in StepOne Plus (Applied Biosystems), using KicqStart Kit (Sigma Aldrich, Merck KGaA, Darmstadt, Germany) with specific primers (sequences in Table S5). All the procedures were conducted according to the manufacturer’s protocol. The relative expressions were calculated using the 2−ΔΔCt method [25 (link)]. The control transcripts used for 2−ΔΔCt were ATP5F1 and RPLP2 as a housekeeper for the mRNA [16 (link),23 (link)].

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Arenas G.A., Valenzuela J.G., Peñaloza E., Paz A.A., Iturriaga R., Saez C.G, & Krause B.J. (2023). Transcriptional Profiling of Human Endothelial Cells Unveils PIEZO1 and Mechanosensitive Gene Regulation by Prooxidant and Inflammatory Inputs. Antioxidants, 12(10), 1874.

Publication 2023

OneScript cDNA Synthesis Kit StepOne Plus KicqStart Kit

Cdna Mrna Primers Quantitative real time pcr Synthesis

Corresponding Organization : University of O'Higgins

Other organizations : Pontificia Universidad Católica de Chile, University of Chile, University of Antofagasta

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Levels of mRNA

control variables

ATP5F1 as a housekeeper for the mRNA
RPLP2 as a housekeeper for the mRNA

controls

Positive control: Not specified
Negative control: Not specified

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