Calibration of denaturing agarose gels for molecular weight estimation of DNA: Size determination of the single‐stranded genomes of parvoviruses

Vertical slab gel electrophoresis of DNA with CH₃HgOH-containing agarose produces sharp bands whose mobilities are suitable for size estimation of single-stranded DNA containing 600-20 000 bases. The relationship of electrophoretic mobility to size of DNA over this range is a smooth, S-shaped function, and an empirical model was developed to express the relationship. The model involves terms in squared and reciprocal mobilities, and produced excellent fit of known standard markers to measured mobilities. It was used to estimate the sizes of six parvovirus DNAs: Kilham rat virus (KRV), H-1, LuIII, and minute virus of mice (MVM) DNAs had molecular weights of 1.66 - 1.70 x 10⁶, while the molecular weight of bovine parvovirus (BPV) DNA was 1.84 x 10⁶ and that of adenoassociated virus (AAV) DNA was 1.52 x 10⁶.