Protocol detail

RNA Extraction and Quantitative RT-PCR

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RNA from cells or tissue powder was isolated using the Roti®-Quick-Kit (Carl Roth). RNA extracted from tissue powder was purified using the Peqlab-Gold Total RNA-Kit (12-6834-01, Peqlab). Reverse transcription and quantitative RT-PCR using the SYBR Green LC480 system (Roche) was conducted as previously described^{41 (link)}. Primers are listed in Supplementary Table 2 and RPL19 or HPRT served as housekeeping genes for relative gene expression analysis.

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Welk V., Meul T., Lukas C., Kammerl I.E., Mulay S.R., Schamberger A.C., Semren N., Fernandez I.E., Anders H.J., Günther A., Behr J., Eickelberg O., Korfei M, & Meiners S. (2019). Proteasome activator PA200 regulates myofibroblast differentiation. Scientific Reports, 9, 15224.

Publication 2019

Roti®-Quick-Kit Peqlab-Gold Total RNA-Kit SYBR Green LC480 system

Cells Gene expression analysis Gold Housekeeping genes Powder Primers Reverse transcription Rt pcr Sybr green Tissue

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Variable analysis

independent variables

RNA isolation method (Roti®-Quick-Kit, Peqlab-Gold Total RNA-Kit)

dependent variables

Gene expression levels measured by quantitative RT-PCR using the SYBR Green LC480 system

control variables

Housekeeping genes (RPL19 or HPRT) used for relative gene expression analysis

controls

No positive or negative controls explicitly mentioned

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