Quantitative RT-PCR Analysis of CCL26

Total RNA was extracted from prepared cultured cells with TRIzol reagent (Invitrogen) and cDNA was synthesized according to the manufacturer's protocol (Roche). Quantitative real-time PCR (qRT-PCR) was performed using a Light Cycler 480 Probe Master System (Roche), and PCR-specific amplification was conducted using the LightCycler^® Nano (Roche) as described previously^{28 (link)}. The relative expression of CCL26 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was calculated using the 2-(ΔΔCt) method method. The primers and probe kits ofCCL26 and GAPDH were obtained from Applied Biosystems (Nagoya, Japan).

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Kawano M., Iwasaki T., Itonaga I., Kubota Y., Tanaka K, & Tsumura H. (2021). Analysis of the signal cross talk via CCL26 in the tumor microenvironment in osteosarcoma. Scientific Reports, 11, 18099.

Publication 2021

TRIzol reagent Light Cycler 480 Probe Master System LightCycler® Nano

Ccl26 Cdna Cultured cells Glyceraldehyde 3 phosphate dehydrogenase Light Primers Quantitative real time pcr Trizol

Corresponding Organization :

Other organizations : Oita University

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Variable analysis

independent variables

TRIzol reagent (Invitrogen)
CDNA synthesis protocol (Roche)
Quantitative real-time PCR (qRT-PCR) using Light Cycler 480 Probe Master System (Roche)
PCR-specific amplification using LightCycler® Nano (Roche)

dependent variables

Relative expression of CCL26
Relative expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH)

control variables

Prepared cultured cells

controls

Positive control: Not specified
Negative control: Not specified

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