Illumina Sequencing of MPECs Genome

MPECs were sequenced using Illumina HiSeq TM2000 platform at the Novogene Bioinformatics (Beijing, China). For Illumina sequencing, at least 1 μg genomic DNA was used for each strain in sequencing library construction. DNA samples were sheared into 400–500 bp fragments using a Covaris M220 Focused Acoustic Shearer following manufacture’s protocol. Illumina sequencing libraries were prepared from the sheared fragments using the NextFlex^TM Rapid DNA-Seq Kit. The prepared libraries then were used for paired-end Illumina sequencing (2 × 150 bp) on an Illumina HiSeq TM2000 machine. The raw data generated from Illumina platform were submitted to Enterobase¹, and sequence types were automatically assigned according to the Achtman scheme. A minimum spanning tree was generated using GrapeTree software to analyze the distribution of sequence type of MPECs (Zhou et al., 2018 (link)) characterized by multilocus sequence typing (MLST) analysis.