RNA gel electrophoresis

The RNA of the 3 samples in duplicates (E.coli K1 treated with 0.5 μg/ml AgNPs-HDN, E.coli K1 treated with 0.5 μg/ml AgNPs, and untreated bacteria) were run on a gel. Briefly, the gel was prepared using 1–1.2% (1X) TEA buffer, heated with mixing until dissolved and 1 μl of gel dye was added while mixing. The gel was poured into a gel tray and allowed to solidify. The tray was transferred into a gel tank and filled with buffer. Next 1 μl of tracking dye was mixed with 5 μl sample. The samples were heated until 95-120 °C for 1 min and placed on ice prior to loading. A 1 kb DNA ladder (5 μl) was pipetted into the first well and the other samples were pipetted in other wells. The gel was run at 100 V for 40 mins and imaged to visualise bands of bacterial RNA 1800 23S and 1200 16S bands. The RNA integrity and concentrations were also checked using Qubit 2.0 fluorometer and the RIN (RNA integrity number) was assessed using Agilent TapeStation 2200 (Table 1).

The criteria of bacterial RNA samples. A0: control, A and B: E. coli K1 treated with AgNPs-HDN, C and D: E. coli K1 treated with AgNPs, E and F: untreated E. coli K1