Protocol detail

Quantitative Gene Expression Analysis

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Total RNA was extracted with an appropriate RNA Extraction Kit (Promega, United States) and reverse-transcribed employing random primers and a Reverse Transcription Kit (Promega, United States). Then, the expression levels of target RNAs were evaluated using a Step One Plus Real-Time PCR System and SYBR Green Master Mix (7300R, ABI, United States). GAPDH was employed as an endogenous control, and the fold change was computed using the 2^−ΔΔCT technique (Mo et al., 2021 (link)). Primer sequences were designed by Sangon Biotech (Table 1).

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Chen N.N., Ma X.D., Miao Z., Zhang X.M., Han B.Y., Almaamari A.A., Huang J.M., Chen X.Y., Liu Y.J, & Su S.W. (2023). Doxorubicin resistance in breast cancer is mediated via the activation of FABP5/PPARγ and CaMKII signaling pathway. Frontiers in Pharmacology, 14, 1150861.

Publication 2023

RNA Extraction Kit Reverse Transcription Kit Step One Plus Real-Time PCR System SYBR Green Master Mix

Gapdh Primer Promega Reverse transcription Rnas expression Step one Sybr green

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression levels of target RNAs

control variables

GAPDH as an endogenous control

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

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