Quantifying Gene Expression via RT-qPCR

Total RNA was isolated using Trizol reagent according to manufacturer instructions (Invitrogen). Complimentary DNA was obtained using High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). Quantitative real-time PCR (QPCR) was performed using Maxima SYBR Green qPCR Master Mixes (Thermo Scientific) as described previously [22 (link)]. Analysis for each gene was carried out three times and normalized to β-actin housekeeping gene expression. Primer information is provided in Supplementary Information.

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Lin Y.W., Park S.W., Lin Y.L., Burton F.H, & Wei L.N. (2019). Cellular retinoic acid binding protein 1 protects mice from high fat diet-induced obesity by decreasing adipocyte hypertrophy. International journal of obesity (2005).

Publication 2019

Trizol reagent High-Capacity cDNA Reverse Transcription Kit Maxima SYBR Green qPCR Master Mixes

Actin Cdna Gene Housekeeping gene Primer Quantitative real time pcr Reverse transcription Sybr green Trizol

Corresponding Organization :

Other organizations : University of Minnesota Medical Center

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Gene expression levels

control variables

β-actin housekeeping gene expression

controls

None specified
None specified

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