Measuring miRNA and mRNA Levels via SYBR Green qPCR

Total RNA was extracted from cultured cells using TRIzol reagent (Invitrogen). The miRNA and mRNA levels were measured using a SYBR Green qPCR assay (TaKaRa, Dalian, China) following the methods described before^{34 (link)}. The expression of GAPDH (reference for mRNA determination) or RNU6B (reference for miRNA determination) served as an endogenous control. The sequences of the PCR primers are presented in Supplementary Table S1. The 2^−ΔCT method was applied for data processing.

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Luo M., Huang P., Pan Y., Zhu Z., Zhou R., Yang Z, & Wang C. (2021). Weighted gene coexpression network and experimental analyses identify lncRNA SPRR2C as a regulator of the IL-22-stimulated HaCaT cell phenotype through the miR-330/STAT1/S100A7 axis. Cell Death & Disease, 12(1), 86.

Publication 2021

TRIzol reagent SYBR Green qPCR assay

Assay Cultured cells Gapdh Mirna Mrna Primers Sybr green Trizol

Corresponding Organization :

Other organizations : Hunan University, Hunan University of Traditional Chinese Medicine

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

MiRNA levels
MRNA levels

control variables

GAPDH (reference for mRNA determination)
RNU6B (reference for miRNA determination)

controls

Positive control: Not specified
Negative control: Not specified

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