Quantitative analysis of Numb gene expression

As described previously [19 (link), 20 (link)], mRNA from PC tissues and cell lines was analyzed in a Light Cycler 2.0 with the Light Cycler kit (Takara Bio, Japan). The primers were as follows: Numb-PRRL, 5’-CTTCCAAGCTAATGGCACTG-3’ (sense) and 5’-CTCTTAGACACCTCTTCTAACCA-3’ (antisense); Numb-PRRS, 5’-CAATCTCCTACCTTCCAAGGG-3’ (sense) and 5’-CGGACGCTCTTAGACACCTC-3’ (antisense). The quality of the PCR products was monitored with post-PCR melt-curve analysis. The expression level of these target genes was calculated by the −ΔΔCt method.

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Sheng W., Tang J., Cao R., Shi X., Ma Y, & Dong M. (2022). Numb-PRRL promotes TGF-β1- and EGF-induced epithelial-to-mesenchymal transition in pancreatic cancer. Cell Death & Disease, 13(2), 173.

Publication 2022

Light Cycler 2.0 Light Cycler kit

Cell lines Expression genes Light Mrna Primers Prrs Tissues

Corresponding Organization : China Medical University

Other organizations : Beijing Chaoyang Emergency Medical Center

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Variable analysis

independent variables

Numb-PRRL primer sequence (sense and antisense)
Numb-PRRS primer sequence (sense and antisense)

dependent variables

Expression level of Numb-PRRL and Numb-PRRS target genes

control variables

MRNA from PC tissues and cell lines
Light Cycler 2.0 with the Light Cycler kit (Takara Bio, Japan)
Post-PCR melt-curve analysis to monitor the quality of PCR products
−ΔΔCt method to calculate the expression level of target genes

controls

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

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