Figure 11.

Liquid hybridization on the immobilized membrane using slot blot could be an alternative to acrylamide gel. Different concentrations of RNA1 and RNA2 (0.1, 0.5 and 1 ug, respectively) were used for liquid hybridization using 10 pmol of biotinylated RN5S or miR-200c probes per RNA concentration. (a) After overnight Exo-I treatment, hybridized mixtures were transferred to positively charged nylon membranes using slot blot, as per manufacturer instructions. After transfer, the UV-crosslinked membrane was incubated with HRP-conjugated streptavidin followed by 5S RNA or miR-200c signal detection using an X-ray. (b) Quantitative analysis of 5S RNA and miR-200c expression levels after X-ray detection. Image quantification was performed using GelQuant.NET software. Relative % intensities were measured for each RNA and plotted against the corresponding amount of RNA.