Ip plate

After total RNA was extracted using an RNeasy Kit according to the manufacturer’s instructions, a northern blot analysis was performed as described previously^{32 (link)}. Briefly, the quantified total RNA samples (2.5 μg) were loaded onto denaturing agarose gels including 0.25 M formaldehyde, separated by gel electrophoresis, and then stained with ethidium bromide to visualize 23S and 16S rRNA. The RNA bands were transferred to nylon membranes (Schleicher and Schuell, Germany) using a TurboBlotter (Schleicher and Schuell, Germany). The membrane was hybridized with a specific ³²P-labeled probe (Takara, Japan) based on PCR amplification with each primer pair. Autoradiography was performed using an IP plate (Fujifilm, Japan) and a Multiplex Bio-Imaging System (Fujifilm, Japan).

Pseudomonas putida KT2440 cells were grown overnight in LB medium and then diluted 100 fold. Appropriate dilutions of exponentially growing cells were spread on LB plates containing indole, Amp, or both indole and Amp. After 36 h of incubation at 30°C, cells were collected from the plates, and total RNA was isolated using an RNeasy Mini Kit according to the manufacturer’s instructions. Northern blot analysis was then performed as described previously (Kim and Park, 2013 (link)). Samples of total RNA (2.5 μg) were loaded onto denaturing agarose gels containing 0.25 M formaldehyde, separated, and then stained with ethidium bromide to visualize 23S and 16S rRNA. The fractionated RNA was transferred to nylon membranes (Schleicher and Schuell, Germany) using a TurboBlotter (Schleicher and Schuell, Germany). The amount of mRNA was determined by hybridizing the membrane with a specific ³²P-labeled probe (Takara, Japan), prepared by PCR amplification with the respective primer pairs. Autoradiography was conducted using an IP plate (Fujifilm, Japan) and Multiplex Bio-Imaging System (Fujifilm, Japan).