Gene Expression Analysis via qPCR

Preparation of cell and tissue extracts, and gene expression analysis, followed previously published methods [11 (link), 30 (link)]. In brief, Sybr green (Biorad) reactions were prepared as a master mix with 50 ng of cDNA/10 μl/well aliquoted into prearrayed and validated (per MIQE guidelines) BioRad pathway finder plates (Epilepsy M384; PrimePCR^TM SABioscience) [31 ]. A BioRad CFX384 real-time PCR with BioRad CFX manager v3.0 software was employed for data acquisition, normalization, and statistical analysis.

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Vogel K.R., Ainslie G.R., Jansen E.E., Salomons G.S., Roullet J.B, & Gibson K.M. (2017). In vitro modeling of experimental succinic semialdehyde dehydrogenase deficiency (SSADHD) using brain-derived neural stem cells. PLoS ONE, 12(10), e0186919.

Publication 2017

Sybr green CFX384 real-time PCR CFX manager v3.0 software

Cdna Cell Epilepsy Gene expression analysis Real time pcr Sybr green Tissue extracts

Corresponding Organization : Amsterdam Neuroscience

Other organizations : Washington State University Spokane

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Variable analysis

independent variables

Preparation of cell and tissue extracts
Gene expression analysis

dependent variables

Gene expression levels

control variables

50 ng of cDNA/10 μl/well aliquoted into prearrayed and validated (per MIQE guidelines) BioRad pathway finder plates (Epilepsy M384; PrimePCR™ SABioscience)
BioRad CFX384 real-time PCR with BioRad CFX manager v3.0 software used for data acquisition, normalization, and statistical analysis

controls

Positive control: Not specified
Negative control: Not specified

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