GSM9060924: HS11286, ΔcpxR, biol rep1; Klebsiella pneumoniae; RNA-Seq
Total bacterial RNA was extracted using the Trizol method, with RNA quality and integrity assessed by agarose gel electrophoresis, NanoDrop spectrophotometer, and Agilent 2100 Bioanalyzer. Ribosomal RNA (rRNA) was removed from total RNA using the Ribo-Zero rRNA Removal Kit (Illumina). First-strand cDNA was synthesized using random oligonucleotides and SuperScript III, followed by second-strand cDNA synthesis employing DNA Polymerase I and RNase H, where dUTP replaced dTTP. Library quality was verified using Agilent 2100, DNA concentration measured with PicoGreen, effective concentration confirmed by qPCR.