WGS of Mycobacterium bovis from bovine lung, China, 2021
Genomic DNA was extracted from the samples using the SDS or STE method. DNA purity and integrity were assessed by agarose gel electrophoresis, and quantification was performed with Qubit. DNA samples that passed electrophoresis quality checks were randomly sheared to ~350 bp fragments using a Covaris ultrasonicator. The resulting DNA fragments were processed with the NEBNext Ultra DNA Library Prep Kit for Illumina (NEB, USA), completing end repair, A-tailing, adapter ligation, purification, and PCR amplification to prepare the sequencing libraries. After library construction, the libraries were first quantified with a Qubit 2.0 and diluted to 2 ng/l. The insert size was then assessed with an Agilent 2100, and, once meeting expectations, the effective library concentration was accurately quantified by qPCR to ensure library quality. After the libraries passed quality control, different libraries were pooled according to their effective concentrations and target data output, and sequenced on an Illumina NovaSeq using PE150.