Protocol detail

Quantifying Gene Expression Using qRT-PCR

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As previously described, total RNA isolation, first-strand cDNA synthesis, and qRT-PCR were performed (30 (link)). The primers were listed in Supplementary Table S1. The transcriptional gene expression was performed with SYBR Green Supermix (Qiagen, Hilden, German) by the qRT-PCR System (StepOnePlus, Applied Biosystems, Waltham, MA, USA). The relative transcriptional level of SCL27A6 was normalized to β-actin mRNA expression and calculated using the 2^−ΔΔCt method (31 (link)).

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Zhong X., Yang Y., Li B., Liang P., Huang Y., Zheng Q., Wang Y., Xiao X., Mo Y., Zhang Z., Zhou X., Huang G, & Zhao W. (2022). Downregulation of SLC27A6 by DNA Hypermethylation Promotes Proliferation but Suppresses Metastasis of Nasopharyngeal Carcinoma Through Modulating Lipid Metabolism. Frontiers in Oncology, 11, 780410.

Publication 2021

SYBR Green Supermix

Actin Cdna Gene expression Isolation Mrna Primers Sybr green Synthesis Transcriptional

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Transcriptional gene expression of SCL27A6

control variables

β-actin mRNA expression

controls

Positive control: Not specified
Negative control: Not specified

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