Quantitative Real-Time PCR for Gene Expression

Quantitative real time-PCR (qRT-PCR) was carried out using DNA Master SYBR Green I mix (Applied Biosystems), as we previously described (Villasante et al., 2014 (link)) mRNA expression levels were quantified applying the ΔCt method, ΔCt = (Ct of gene of interest—Ct of GAPDH). qRT-PCR primer sequences that were obtained from the PrimerBank data base¹ are listed in Table 1. Primer sequences for CD206, CD163, CCR7, CD80, and TNFα were obtained from Spiller et al. (2014) .

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Villasante A., Robinson S.T., Cohen A.R., Lock R., Guo X.E, & Vunjak-Novakovic G. (2021). Human Serum Enhances Biomimicry of Engineered Tissue Models of Bone and Cancer. Frontiers in Bioengineering and Biotechnology, 9, 658472.

Publication 2021

DNA Master SYBR Green I mix

Cd163 Gapdh Gene Mrna Primer Quantitative real time pcr Sybr green i Tnfα

Corresponding Organization : Universitat de Barcelona

Other organizations : Drexel University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

MRNA expression levels of genes of interest

control variables

GAPDH expression level (used as reference gene)

controls

Positive controls: None mentioned
Negative controls: None mentioned

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