Rnase away

Microdissections were performed on a cold plate (Weinkauf Medizintechnik, Forchheim, Germany) set at -20°C as previously described (Brunner et al., 2014 (link)). Care was taken to thoroughly clean the working area and dissection instruments with RNase AWAY (Carl Roth, Karlsruhe, Germany) before each dissection. Hypothalamic brain areas (Bregma +0.38 to -2.92) were collected in MagnaLyser bead tubes (catalog number: 03358 941 001, Roche Diagnostics, Rotkreuz, Switzerland) filled with Precellys beads (Peqlab, Erlangen, Germany) and stored at -70°C until RNA extraction.

The brain microdissection of the mPFC was performed as described previously^{66 (link)}. After cleaning the working area and dissection instruments with RNase AWAY (Carl Roth, Karlsruhe, Germany), the brains were cut into 1 mm thick slices at −20 °C in the cryostat. Then the mPFC (Bregma, +3.20 to −0.22) was micro-dissected under a stereomicroscope on a cold plate (Weinkauf Medizintechnik, Forchheim, Germany) at −20 °C. The dissected mPFC was collected in micro packaging vials filled with Precellys beads (Peqlab, Erlangen, Germany) and stored at −70 °C until RNA extraction.

If not otherwise stated, chemical reagents were purchased from Sigma-Aldrich (Vienna, Austria). Denatured ethanol (EtOH) was purchased from Carl Roth (Vienna, Austria), deionized formamide was purchased from PanReac AppliChem (Darmstadt, Germany), EtOH absolute was purchased from AustrAlco (Spillern, Austria), and UltraPure salmon sperm DNA solution was purchased from Thermo Fisher Scientific (Vienna, Austria). Cell culture media, media supplements and antibiotics were purchased from Thermo Fisher Scientific. All buffers were prepared using ultrapure water (Milli-Q, 18.2 MΩ·cm at 25 °C, Merck Millipore, Vienna, Austria) and filtered through sterile 0.22 µm polyvinylidene fluoride (PVDF) syringe filters (Carl Roth, Austria). Surfaces and instruments were wiped with the surface decontaminant RNase Away (Carl Roth, Austria) prior to work.