Measuring Gene Expression Using Real-Time PCR

Applied Biosystems QuantStudio 7 Flex Real-Time PCR was used to measure gene expression using Taqman Gene Expression Assay Kits (Thermo Fisher). Seven different housekeeping genes were tested using Normfinder and the three most stable genes between the three cell lines were used to normalise gene expression across samples as previously described [18 (link)]. Relative fold change in gene expression was calculated using the 2^−ΔΔCT method and represented relative to a control sample set to 1 for each experiment. Data presented is from three independent experiments displayed as mean +/− SEM.

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Mahdi A.F., Malacrida B., Nolan J., McCumiskey M.E., Merrigan A.B., Lal A., Tormey S., Lowery A.J., McGourty K, & Kiely P.A. (2020). Expression of Annexin A2 Promotes Cancer Progression in Estrogen Receptor Negative Breast Cancers. Cells, 9(7), 1582.

Publication 2020

QuantStudio 7 Flex Real-Time PCR Taqman Gene Expression Assay Kits

Assay Cell lines Gene expression Genes Housekeeping genes Real time pcr

Corresponding Organization : University of Limerick

Other organizations : University Hospital Limerick, Ollscoil na Gaillimhe – University of Galway

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Gene expression

control variables

Seven different housekeeping genes were tested using Normfinder and the three most stable genes between the three cell lines were used to normalise gene expression across samples as previously described [18 (link)]

controls

Positive control: None mentioned
Negative control: None mentioned

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