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To confirm the results shown by gel retardation analysis this second technique was chosen.
To determine if the lipid-polymer hybrid NPs could complex negatively charged siRNA, a gel retardation assay was conducted after incubating the nanoparticles with siRNA (100 nM) at different N/P ratios.
(c) Gel retardation assay of siRNA complexed with nanoparticles at different N/P ratios.
Gel retardation studies were used to determine DNA binding ratio of SLNs.
Common techniques for this include hybridization of a specific probe to the PCR product after Southern blotting or by liquid hybridization followed by gel retardation analysis (12,14,17).
One aliquot was analyzed by liquid hybridization followed by gel retardation. The second PCR aliquot was analyzed by a different operator in a blinded fashion directly by microchip electrophoresis using an instrument built in-house from commercially available components.
Agarose Gel Retardation. SS-PEI/DNA complexes were prepared by adding a HEPES buffer solution (20 mM, pH 7.4) of SS-PeI (10 [micro]L, varying concentrations) to a HEPES solution of DNA (10 [micro]L, 80 [micro]g/mL), followed by vortexing for 5 s, and the dispersions were incubated for 30 min at room temperature.
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- gel electrophoresis
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- gel-filtration chromatography, gel-permeation chromatography
- gel-filtration chromatography, gel-permeation chromatography
- gel-filtration chromatography, gel-permeation chromatography
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