After the retrieval, tissues were transferred in antibiotic solution made up with Gentamicin, Vancomycin and Meropenem and maintained at + 4 °C following the validated FBTV internal procedures (Serafini et al. 2016 (link); Montagner et al. 2018 (link); Paolin et al. 2018 (link)).
Before the analysis, tissues were kept at + 4 °C.
The tissues were manually dissected into few sections of approximately 2 mm and treated over night with MagNA Pure Bacteria Lysis Buffer (Roche). Detection of SARS-CoV-2 RNA was performed by an in-house real-time RT–PCR method, which was developed according the protocol and the primers and probes designed by CDC that targeted the gene encoding nucleocapsid (N2) of SARS-CoV-2 and RNA-dependent RNA polymerase. Real-time RT–PCR assays were performed in a final volume of 25 μl, containing 5 μl of purified nucleic acids, using One Step Real Time kit (Thermo Fisher Scientific) and run on ABI 7900HT Fast Sequence Detection Systems (Thermo Fisher Scientific).
Before the analysis, tissues were kept at + 4 °C.
The tissues were manually dissected into few sections of approximately 2 mm and treated over night with MagNA Pure Bacteria Lysis Buffer (Roche). Detection of SARS-CoV-2 RNA was performed by an in-house real-time RT–PCR method, which was developed according the protocol and the primers and probes designed by CDC that targeted the gene encoding nucleocapsid (N2) of SARS-CoV-2 and RNA-dependent RNA polymerase. Real-time RT–PCR assays were performed in a final volume of 25 μl, containing 5 μl of purified nucleic acids, using One Step Real Time kit (Thermo Fisher Scientific) and run on ABI 7900HT Fast Sequence Detection Systems (Thermo Fisher Scientific).
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