GSM9789213: Listeria monocytogenes F2365 delta-buk mutant, biological replicate 2; Listeria monocytogenes; RNA-Seq
Total RNA was extracted from two independent biological replicates per condition using the FastRNA spin kit for microbes with the FastPrep-24 instrument (MP Biomedicals, Santa Ana, CA). Genomic DNA contamination was removed by on-column DNase treatment (RNase-free DNase set, Qiagen). RNA quantity and purity were assessed by NanoDrop ND-1000 spectrophotometry. Ribosomal RNAs were depleted using the Ribo-Zero magnetic kit for Gram-positive bacteria (Epicentre), and enriched mRNA was fragmented prior to library preparation. RNA-seq libraries were constructed and sequenced by Novogene Co. Fragmented, rRNA-depleted RNA was reverse transcribed into cDNA using random hexamer primers. Libraries were prepared following Novogene standard strand-specific library preparation protocols. Sequencing was performed on an Illumina NovaSeq 6000 platform using 150-bp paired-end (PE150) strategy.