Determination of agarose gel pore size: Absorbance measurements vis a vis other techniques

Abstract

The absorbance measurements in the wavelength range 700 nm to 800 nm were used to probe the agarose gel topology evolution and extract the pore size of the trapped solvent. By following the changes in absorbance and pore size, the gelation process could be clearly divided into three stages - induction stage, gelation stage and pseudo-equilibrium stage. The gelation mechanism is explained as a nucleation and growth process. Following the kinetics of gelation using dynamic light scattering is complicated by multiple scattering (for high concentrations) and large fluctuations in intensity and relaxation time. Comparatively, scanning the absorption spectrum is fast and the method is suitable for a wide range of concentrations and setting temperatures. Pore size determination using absorbance is a fast and non-invasive method when compared to the DNA electrophoresis measurements, which extend over several hours and use probe diffusion.