Computational protocol: Region-Specific Differences in Amyloid Precursor Protein Expression in the Mouse Hippocampus

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Protocol publication

[…] cDNA was amplified using TaqMan® Fast Universal PCR Master Mix (Applied Biosystems) and the StepOnePlus Real-Time PCR System (Applied Biosystems). PCR products were checked on Agilent DNA 1000 Chips (Agilent Technologies) with the Agilent 2100 Bioanalyzer system to verify product specificity and amplicon size. Quantification of the gene expression of candidate reference genes was carried out using SYBR® GreenERTM qPCR Supermix Universal (Invitrogen, Waltham, MA, USA) following the manufacturer’s recommendations. Primer efficiencies and quantification cycle (Cq) values were calculated using LinRegPCR Software (Tuomi et al., ). To determine the most stable reference genes and the minimum number for accurate normalization, NormFinder (Andersen et al., ) and geNorm (Vandesompele et al., ) were used according to the developer’s manuals. qPCR data were tested for statistical significance using one-way ANOVA followed by Bonferroni post hoc test to correct for multiple comparisons, *p ≤ 0.05. […]

Pipeline specifications

Software tools LinRegPCR, NormFinder, geNorm
Application qPCR
Organisms Mus musculus