Brain Region Transcriptome Analysis

The total RNA of homogenized tissue from specific brain regions was isolated using the Trizol^® method according to manufacturer’s instructions (GibcoBRL) and its concentration was determined using the nanodrop ND-1000 spectrophotometer followed by its quality and integrity measurement by electrophoresis on RNA 6000 LabChip^® kit (Agilent 2100 Bioanalyzer). The RNA was transcribed into the cDNA using the iScript cDNA Synthesis Kit (BioRad). The primer pairs used for RT-PCR are the same used previously for quantitative RT-PCR (Vogelgesang et al., 2017 (link)).

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Niebert S., van Belle G.J., Vogelgesang S., Manzke T, & Niebert M. (2017). The Serotonin Receptor Subtype 5b Specifically Interacts with Serotonin Receptor Subtype 1A. Frontiers in Molecular Neuroscience, 10, 299.

Publication 2017

Trizol® method nanodrop ND-1000 spectrophotometer RNA 6000 LabChip® kit iScript cDNA Synthesis Kit

Brain Cdna Electrophoresis Instructions method Primer Rt pcr Synthesis Tissue Trizol

Corresponding Organization : Nanoscale Microscopy and Molecular Physiology of the Brain Cluster of Excellence 171 — DFG Research Center 103

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Variable analysis

independent variables

Brain regions

dependent variables

Total RNA concentration
RNA quality and integrity

control variables

Trizol method for RNA isolation
Nanodrop ND-1000 spectrophotometer for RNA concentration measurement
RNA 6000 LabChip kit (Agilent 2100 Bioanalyzer) for RNA quality and integrity measurement
IScript cDNA Synthesis Kit (BioRad) for cDNA synthesis
Primer pairs used for RT-PCR

controls

Positive control: None mentioned
Negative control: None mentioned

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