microarray crna synthesis, hybridisation and plate-reading  (Geneservice ltd)

Journal: mBio

Article Title: Physiological roles of an Acinetobacter -specific σ factor

doi: 10.1128/mbio.00968-25

Figure Lengend Snippet: SigAb mitigates and responds to copper stress. (A) Heatmap of heavy metal resistance genes significantly upregulated (log 2 FC > 1, FDR < 0.05, T = 10 min) in RNA-seq time course experiment of sigAb overexpression strain compared to empty vector control. Operons are denoted to the right, with arrows indicating the direction of transcription. (B) Growth curves plotted as OD 600 over time (h) of CRISPRi sigAb knockdown (KD) strain and non-targeting (NT) control in rich defined medium with 250 µg/mL CuSO 4 stress ( n = 3). Data are represented as mean ± s.d. for NT control with no stress (red), sigAb KD with no stress (blue), NT control with copper stress (green), and sigAb KD with copper stress (purple). (C) sigAb KD growth defects in metal stresses graphed as area under the curve normalized to NT control ( n = 2–9). Data are represented as the mean ± s.d., and significance was calculated with a two-tailed Student’s t -test ( P < 0.05). Bars without asterisks are not significantly different from the control. (D) SigAb induction curves plotted as P sigAb activity (mRFP fluorescence) versus cell density (OD 600 ) for metal and antibiotic stress conditions using Biolog Phenotype Microarray PM13.

Article Snippet: To expand our phenotyping to additional conditions (e.g., manganese, cobalt), we tested the growth of the sigAb knockdown in Biolog Phenotype Microarray (PM) plates ( ; ).

Techniques: RNA Sequencing, Over Expression, Plasmid Preparation, Control, Knockdown, Two Tailed Test, Activity Assay, Fluorescence, Microarray