Real-time RT-qPCR for Adipogenic Gene Expression

Real-time RT-qPCR was performed as described previously [11 (link)]. Total RNA was isolated using the TRIzol Reagent (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). RNA (500 ng) was reverse transcribed with the High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor (Applied Biosystems, Thermo Fisher Scientific) according to the manufacturer’s instructions. The mRNA levels of adipogenic-related genes were measured with Single Tube TaqMan Gene Expression Assays (Life Technologies, Thermo Fisher Scientific) (Supplementary Table S8). The levels of gene expression were quantified relative to the level of Hprt1.

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Walendzik K., Bukowska J., Kopcewicz M., Machcinska S., Gimble J.M, & Gawronska-Kozak B. (2020). Age, Diet and Epidermal Signaling Modulate Dermal Fibroblasts’ Adipogenic Potential. International Journal of Molecular Sciences, 21(23), 8955.

Publication 2020

TRIzol Reagent High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor Single Tube TaqMan Gene Expression Assays

Adipogenic Assays Cdna Gene expression Genes Mrna Reverse transcription Rnase Trizol

Corresponding Organization : Polish Academy of Sciences

Other organizations : LaCell (United States), Tulane University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

MRNA levels of adipogenic-related genes

control variables

The levels of gene expression were quantified relative to the level of Hprt1

controls

No positive or negative controls were explicitly mentioned.

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