Fasti001 1 favorprep stool dna isolation mini kit

Before processing, frozen samples were thawed at room temperature. From this step, all samples were processed in the same way. Direct DNA extraction was carried out using a commercial kit (FASTI001-1 FavorPrep Stool DNA Isolation Mini Kit, Favorgen-Europe, Vienna) following the manufacturer’s specifications (elution volume of 200 µL), coupled to a specific SYBRGreen real-time PCR assay for quantitative detection of the mcr-1 gene (qPCR), described previously [49 (link)] and further validated in our lab [13 (link)]. Samples were considered positive if their quantitative DNA values were > 1.00 × 10² fg/µL, equivalent to 1.58 × 10³mcr-1 copies/mg caecal content.

Direct DNA extraction from chick caecal samples was carried out using a commercial kit (FASTI001-1 FavorPrep Stool DNA Isolation Mini Kit, Favorgen-Europe, Vienna, Austria) following the manufacturer’s specifications (elution volume of 200 µL), coupled with a specific SYBRGreen (Thermo Fisher Scientific, Vilnius) real-time PCR assay for quantitative detection of the mcr-1 gene (qPCR), as described previously by Li J et al. [28 (link)] and further validated in our previous work [29 (link)]. Samples were considered positive if quantitative values were >1.00 × 10² fg/µL (equivalent to 1.58 × 10³ copies/mg caecal content).