ATCC_BT with RA-CH-2[ere(D)] gDNA isolate No.8
The genomic DNA (gDNA) of Riemerella anatipestifer RA-CH-2, which contains the tetracycline resistance gene tet(X) and erythromycin resistance gene ere(D), served as the donor, with ATCC_BT [contains blaRATA-2 and tet(X) genes] as the recipient strain. Natural transformants were obtained through selection with sub-inhibitory concentrations of erythromycin. First, magnetic beads were used to screen large fragments of DNA; then library construction is performed: a) DNA (~1ug) repair and end-prep; b) The repaired DNA was purified and recovered using magnetic beads method. Qubit fluorometer was used to quantify DNA and calculate the recovery rate; c) Each sample was added with the corresponding barcode and the barcode-linked product was purified by magnetic beads; d) Calculated the pooling ratio for multiple barcode connection products according to the data volume requirements, and pooling according to the ratio; e) Linker connection, so that each DNA fragment has a unique linker sequence that can be identified; f) Connected DNA was purified by magnetic bead method, Qubit fluorometer was used to quantify DNA and calculate the recovery rate; Finally, starting a sequencing run.