Animals were euthanised by live decapitation with no anaesthesia after 1 or 30D of forced abstinence. Whole brains were quickly flash frozen. The NAc core was dissected by gross microdissection on dry ice. Total RNA was extracted from NAc core tissue using the Mirvana Paris Protein & RNA Isolation System (Thermo Fisher Scientific, Waltham, MA) according to manufacturer's instructions as previously described (Sillivan et al., 2019 (link)). The RNA was suspended in RNase free water, and the concentration of the RNA was measured using the Qubit HS Assay Kit (Life Technologies Corporation of Thermo Fisher Scientific, Frederick, Maryland). One animal was excluded from the molecular analysis due to low RNA yield.
Protocol detail
Extraction of NAc Core RNA
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Anaesthesia
Animal
Assay
Brains
Decapitation
Dry ice
Frozen
Isolation
Microdissection
Protein
Rnase
Tissue
Corresponding Organization : Temple University
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Variable analysis
independent variables
- Duration of forced abstinence (1 day or 30 days)
- Total RNA extracted from the NAc core tissue
- Method of euthanasia (live decapitation with no anaesthesia)
- Brain dissection method (gross microdissection on dry ice)
- RNA extraction method (Mirvana Paris Protein & RNA Isolation System)
- RNA concentration measurement method (Qubit HS Assay Kit)
- Positive control: Not specified
- Negative control: Not specified
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