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For histology, blood sampling, and RNA/protein extraction animals were sacrificed at day 14 corresponding to day 12 after surgery (Fig. 3A).
At the day of tissue extraction animals were sacrificed by cervical dislocation and five tissue samples were collected from each animal.
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For RNA and protein extractions, animals were sacrificed by cervical dislocation without anesthesia.
To determine whether the cytoplasmic inclusions contained insoluble aggregates of FUS, we undertook sequential extraction of animals with RIPA and then with urea buffer.
Four days after venom extraction, one animal of each species was euthanized under pentobarbital anesthesia (>100 mg/kg), in accordance with OIST animal care protocols.
Microbial production of chondroitin and chondroitin-like polysaccharides from renewable feedstock is a promising and sustainable alternative to extraction from animal tissues.
The latter kit was used as a gold standard, as a literature review indicated that this kit was used in almost all recent studies (2005 2008) dealing with DNA extraction from animal and human stools.
For DNA extraction the animal was thawed, weighed and Prepman Ultra (Applied Biosystems) was added directly to the tube at the following rates: ≤0.20 g, 450 500 µL; 0.21 0.30 g, 500 750 µl; >0.31 g, 900 µL.
The extraction from animal mucosal tissues may also introduce risks of pathogen contamination.
After venom extraction, the animal was allowed to recover for four days while transcription levels reached their maxima [ 87].
As of yet, we have no evidence that the E. coli-derived 4 × T β4 has similar binding characteristics equal to the commercial T β4, which is generated by chemosynthesis or extraction of animal thymuses.
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