DNA-Seq of Klebsiella pneumoniae: patients
Genomic DNA was extracted from plant material using the Tiangen DP305 Plant Genomic DNA Kit. DNA concentration was quantified using a NanoDrop 2000 spectrophotometer and quality was verified by agarose gel electrophoresis, ensuring a minimum input of >50 ng for library construction. Sequencing libraries were prepared using the TruePrep DNA Library Prep Kit V2 for Illumina (Vazyme). This kit employs a single-tube transposase reaction to simultaneously fragment the genomic DNA and ligate Illumina-compatible adapters in one step. A subsequent limited-cycle PCR amplification was performed to add unique dual indexes (UDIs) to each sample library. Library quality and size distribution were assessed using the QIAxcel Advanced Automatic nucleic acid analyzer. Final library quantification for accurate pooling was performed via qPCR using the KAPA SYBR FAST qPCR Kit. The pooled libraries were sequenced on an Illumina NovaSeq platform, generating 175 base pair paired-end reads.