Quantitative RT-PCR Analysis of Gene Expression

RNA was isolated using TRIzol (Life Technologies). Conversion to cDNA and subsequent qRT-PCR analysis were performed using the High-Capacity cDNA Reverse Transcription Kit and Taqman Gene expression Master Mix (Applied Biosystems). Predesigned Taqman primer/probe mixes were purchased from Integrated DNA Technologies. Data are presented as fold changes in RNA levels compared to control treatment, calculated following the 2^−ΔΔCt method, with GAPDH and Actin Beta serving as two internal control genes [17 (link), 18 (link)].

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Yoshida K., Taga T., Saito M., Suematsu S., Kumanogoh A., Tanaka T., Fujiwara H., Hirata M., Yamagami T., Nakahata T., Hirabayashi T., Yoneda Y., Tanaka K., Wang W.Z., Mori C., Shiota K., Yoshida N, & Kishimoto T. (1996). Targeted disruption of gp130, a common signal transducer for the interleukin 6 family of cytokines, leads to myocardial and hematological disorders.. Proceedings of the National Academy of Sciences of the United States of America, 93(1), 407-411.

Publication 2019

TRIzol Taqman Gene expression Master Mix Taqman primer/probe mixes

Actin beta Cdna Gapdh Gene expression Genes control Primer Reverse transcription Trizol

Corresponding Organization :

Other organizations : Jesse Brown VA Medical Center, University of Illinois at Chicago, Midwestern University

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Variable analysis

independent variables

Treatment (control vs. experimental)

dependent variables

Fold changes in RNA levels

control variables

GAPDH
Actin Beta

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