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For virus isolation, a 10% suspension of brain sample was prepared and injected into flasks of RK13, equine dermal, and Vero-MARU cells (Vero M).
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Plasma and brain samples were prepared by precipitation with n-propanol.
Brain samples were prepared as 20% homogenates in 5% (w/v) glucose prior to dilution (1/100) in EIA buffer.
Brain samples were prepared for electron microscopy as previously outlined in detail [ 34].
The mouse brain samples were prepared using the established techniques described in the literature [ 17].
Human brain samples were prepared as 20% homogenates and equal protein concentrations then prepared for SDS PAGE by addition of SDS PAGE sample buffer and heating to 100°C.
Fresh frozen autopsied human brain samples were prepared as 0.8 1.0 cm fragments for microarray gene expression analysis from frontal cortex dissections, which were validated by quantitative real-time polymerase chain reaction (Torres-Munoz JE, unpublished data, 2006).
Frozen mouse brain samples were prepared as 10% (wt/vol) homogenates in 5% glucose in distilled water in grinding tubes (Bio-Rad, Hercules, CA, USA) by using a TeSeE Precess 48 homogenizer (Bio-Rad) following the manufacturer's instructions.
The blood and brain samples were prepared by spiking 75 μL of the corresponding IS working solutions (carbamazepine, 10 ng/mL) into 15 μL of dialysate followed by vortex mixing.
Test sample was prepared in distilled water.
Each sample was prepared in quadruplicate.
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