Quantitative Real-Time PCR Protocol

RT-qPCR was performed as previously described 11 (link). Total RNA was extracted according to the instructions for Trizol reagent (Invitrogen, Waltham, MA, United States). PrimeScript RT Master Mix [Takara Biotechnology (Dalian) Co., Ltd., Japan] were reverse transcribed into cDNA, and finally qRT-PCR assay was performed according to Premix Ex TaqTM II [Takara Biotechnology (Dalian) Co., Ltd.]. GAPDH was used as an internal reference gene to calibrate relative expression levels. The sequence of primers of genes to be detected is shown in Table S3.

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Lv Z., Wang M., Hou H., Tang G., Xu H., Wang X., Li Y., Wang J, & Liu M. (2023). FOXM1-regulated ZIC2 promotes the malignant phenotype of renal clear cell carcinoma by activating UBE2C/mTOR signaling pathway. International Journal of Biological Sciences, 19(11), 3293-3306.

Publication 2023

Trizol reagent PrimeScript RT Master Mix Premix Ex TaqTM II

Assay Cdna Gapdh Genes Primers Trizol

Corresponding Organization :

Other organizations : Chinese Academy of Medical Sciences & Peking Union Medical College, Xiangya Hospital Central South University, Central South University, Second Xiangya Hospital of Central South University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Relative expression levels of genes

control variables

GAPDH as an internal reference gene to calibrate relative expression levels

controls

No positive or negative controls were explicitly mentioned

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