Real-time qPCR Analysis of Gene Expression

The total RNA of cell lines and tissues were extracted using Trizol reagent (Invitrogen, Carlsbad, CA) and were reverse transcribed to cDNA via one-step RT-PCR kit (TransGen Biotech Co., Ltd, Beijing, China) according to the manufacturer’s instructions. Real-time quantitative polymerase chain reaction was performed in an Applied Biosystems 7500 Detection system with SYBR Green (TransGen Biotech Co., Ltd, Beijing, China). The relative mRNA levels of genes were normalized to peptidylprolyl isomerase A (PPIA) [32 (link)] using the 2^-ΔΔCT method. The primer sequences are given in Additional file 1: Table S2.

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Ma X., Gu L., Li H., Gao Y., Li X., Shen D., Gong H., Li S., Niu S., Zhang Y., Fan Y., Huang Q., Lyu X, & Zhang X. (2015). Hypoxia-induced overexpression of stanniocalcin-1 is associated with the metastasis of early stage clear cell renal cell carcinoma. Journal of Translational Medicine, 13, 56.

Publication 2015

Trizol reagent one-step RT-PCR kit 7500 Detection system SYBR Green

Cdna Cell lines Genes Mrna Peptidylprolyl isomerase Primer Real time polymerase chain reaction Rt pcr Sybr green Tissues Trizol

Corresponding Organization : Chinese PLA General Hospital

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Variable analysis

independent variables

Cell lines and tissues

dependent variables

Relative mRNA levels of genes

control variables

Peptidylprolyl isomerase A (PPIA)

controls

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

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