2.4.4. Euthanasia and PCR analysis
Mice were bled out under gaseous anaesthesia (CO2) via heart puncture. Target organs/tissues were harvested, immediately flushed free of blood by infusion of pre-warmed PBS to avoid contamination with BTs, and they were then thawed and ground up using a Potter-Elvehjem. Organs/tissues DNA extraction and PCR was performed as stated above (Martín-Escolanoet al. , 2018; Paucar et al. , 2019) (Scheme 2 ). Finally, the PCR products were visualised by UV illumination after a 2 % agarose gel electrophoresis for 90 min at 90 V, containing 1:10000 GelRed nucleic acid gel stain.
In addition, spleens and hearts were weighed to evaluate inflammation of this organ in the different groups of mice.