Gel electrophoresis

Agarose gel electrophoresis (1%) was used to assess complex formation between HNTs and siRNA, using mass ratios of HNTs to siRNAs of 100:1, 50:1, 30:1, 20:1, 10:1, and 1:1. The resulting HNTs/siRNA complexes were left for 30 min to form complexes at room temperature, after which the samples were subjected to centrifugation at 5000 rpm for 5 min. The suspensions were reserved for analysis. The pellets were electrophoresed through 1% agarose gels and with ethidium bromide (0.1 μg/ml) at 100 V for 40 min in tris-acetate (TAE) buffer (0.045 M TAE; 0.001 M EDTA); the bands were visualized on an ultraviolet transilluminator.