Quantitative RT-PCR Analysis Protocol

Quantitative real-time (RT) PCR analysis was performed using an established procedure [19 (link)]. Total RNA was extracted using QIAGEN extraction kit and reverse transcription was performed using SuperScript III RNase H (Invitrogen, Waltham, MA, USA). Quantitative RT-PCR was performed using SYBR Green I and a LightCycler (Roche Diagnostics, Castle Hill, Australia). For normalized gene expression, 18S was used as the reference gene in all experiments. Table S1 lists the primers used for RT-PCR analysis.

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Lee S.J., Nam M.J., Lee D.E., Park J.W., Kang B.S., Lee D.S., Lee H.S, & Kwon O.S. (2018). Silibinin Ameliorates O-GlcNAcylation and Inflammation in a Mouse Model of Nonalcoholic Steatohepatitis. International Journal of Molecular Sciences, 19(8), 2165.

Publication 2018

SuperScript III RNase H SYBR Green I LightCycler

Diagnostics Gene Gene expression Primers Quantitative real time pcr analysis Real time pcr analysis Reverse transcription Rnase h Sybr green i

Corresponding Organization : Kyungpook National University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Normalized gene expression

control variables

Total RNA extraction using QIAGEN extraction kit
Reverse transcription using SuperScript III RNase H (Invitrogen, Waltham, MA, USA)
Quantitative RT-PCR using SYBR Green I and a LightCycler (Roche Diagnostics, Castle Hill, Australia)
18S as the reference gene for normalized gene expression

controls

Positive control: None mentioned
Negative control: None mentioned

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