RNA Extraction and Expression Analysis

RNA extraction was conducted by employing TRIzol™ (Invitrogen), following the methods described before [27 (link)] with SYBR Green PCR Master Mix (Qiagen) for mRNA expression level determination and a Hairpin-it miRNAs qPCR kit (GenePharma) for miRNA expression level determination. The expression levels of GAPDH and RNU6B were used as endogenous controls. The primer sequences for PCR were shown in Table 1. Finally, the data were processed using the 2^−ΔΔCt relative expression method. The more method details of real time PCR assay listed in Additional file 1.

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Xiong Y., Zu X., Wang L., Li Y., Chen M., He W, & Qi L. (2021). The VIM-AS1/miR-655/ZEB1 axis modulates bladder cancer cell metastasis by regulating epithelial–mesenchymal transition. Cancer Cell International, 21, 233.

Publication 2021

TRIzol™ SYBR Green PCR Master Mix Hairpin-it miRNAs qPCR kit

Assay Gapdh Mirna Mrna Primer Real time pcr Sybr green Trizol

Corresponding Organization :

Other organizations : Second Xiangya Hospital of Central South University, Central South University, Xiangya Hospital Central South University

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Variable analysis

independent variables

RNA extraction method: TRIzol (Invitrogen)

dependent variables

MRNA expression levels
MiRNA expression levels

control variables

GAPDH and RNU6B expression levels as endogenous controls

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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