SYBR Green qRT-PCR Protocol for Gene Expression

All primers in this study (S1 Table) were designed using Primer 3 (http://frodo.wi.mit.edu/primer3/), and synthesized by Eurofins MWG Operon. SYBR Green based qRT-PCR was performed using ABsolute Blue qPCR SYBR Green ROX Mix (AB-4162, Thermo Scientific), using a Step One Plus Real-Time PCR System (Applied Biosystems). For qRT-PCR data analysis, the fold change in the target gene relative to the GAPDH (glyceraldehyde 3-phosphate dehydrogenase) endogenous control gene was determined by the 2^–Δ(ΔCt) method, as described previously [6 (link), 15 (link), 18 (link)–21 (link)].

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Qin F., Song Y., Zhang Y., Facemire L., Frierson H, & Li H. (2016). Role of CTCF in Regulating SLC45A3-ELK4 Chimeric RNA. PLoS ONE, 11(3), e0150382.

Publication 2016

ABsolute Blue qPCR SYBR Green ROX Mix Step One Plus Real-Time PCR System

Gapdh Gene Glyceraldehyde 3 phosphate dehydrogenase Operon Primer Step one Sybr green

Corresponding Organization : University of Virginia

Other organizations : Zhejiang Academy of Medical Sciences

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Fold change in the target gene relative to the GAPDH (glyceraldehyde 3-phosphate dehydrogenase) endogenous control gene

control variables

GAPDH (glyceraldehyde 3-phosphate dehydrogenase) endogenous control gene

controls

Positive control: None mentioned
Negative control: None mentioned

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