qRT-PCR for miRNA and mRNA Quantification

For miRNA qRT-PCR the miScript SYBR® Green PCR Kit (Qiagen) and for mRNA the SYBR® Green PCR Master Mix (Applied Biosystems®) was used. In both instances, the manufacturer’s instructions were followed. Each cDNA sample was tested in duplicate (as in comparable studies^{24 (link)–26 (link)} and dCT was determined by normalization of the Ct value to the Ct value of the reference small nucleolar RNA SNORD61 (Qiagen) for miRNA or to beta-actin (Invitrogen, Carlsbad, USA) for mRNA. Data were transformed into relative values by calculating: 2−ΔΔCT (ddCT) as previously described^{23 (link)}.

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Ronovsky M., Zambon A., Cicvaric A., Boehm V., Hoesel B., Moser B.A., Yang J., Schmid J.A., Haubensak W.E., Monje F.J, & Pollak D.D. (2019). A role for miR-132 in learned safety. Scientific Reports, 9, 528.

Publication 2019

miScript SYBR® Green PCR Kit SYBR® Green PCR Master Mix SNORD61 beta-actin

Beta actin Cdna Mirna Mrna Small nucleolar rna Sybr green

Corresponding Organization :

Other organizations : Medical University of Vienna, Research Institute of Molecular Pathology

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Variable analysis

independent variables

Unspecified

dependent variables

MiRNA expression levels
MRNA expression levels

control variables

Normalization to the reference small nucleolar RNA SNORD61 for miRNA
Normalization to beta-actin for mRNA

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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