Quantitative RT-PCR Analysis of Gene Expression

Total RNA was extracted with TransZol reagent (TransGen, Cat. No. ET111-01) as previously described.^{86 (link)} Thereafter, total RNA was reverse transcribed using the GoldScipt cDNA Synthesis Kit (Invitrogen, Cat. No. C81401190) and subjected to quantitative RT-PCR analysis in an ABI 7300 Detection System (Applied Biosystems) using the SYBR Green PCR Master Mix (Applied Biosystems, Cat. No. 4344463) according to the manufacturer’s instructions. GAPDH served as reference, and the 2^−ΔΔCT method^{87 (link)} was used to analyze the data. The primers used for qRT-PCR were as listed in Supplementary Information, Table S1.

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Zheng P., Chen Q., Tian X., Qian N., Chai P., Liu B., Hu J., Blackstone C., Zhu D., Teng J, & Chen J. (2018). DNA damage triggers tubular endoplasmic reticulum extension to promote apoptosis by facilitating ER-mitochondria signaling. Cell Research, 28(8), 833-854.

Publication 2018

TransZol reagent ABI 7300 Detection System SYBR Green PCR Master Mix

Cdna Gapdh Primers Rt pcr Sybr green Synthesis

Corresponding Organization :

Other organizations : Peking University, Institute of Biophysics, National Institute of Neurological Disorders and Stroke, National Institutes of Health

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Gene expression levels measured by quantitative RT-PCR

control variables

GAPDH expression as a reference gene

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

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