Abstract
Several polysaccharides were employed as a molecule-sieving additive for the separation of biological macromolecules in capillary electrophoresis (CE). Especially, glucomannan with an average molecular weight of 3x105∼2x106 Da, prepared from the crude polysaccharide of Amorphophallus konjac, exhibited good resolution and reproducibility on separation of biological macromolecules. Under optimized conditions, restriction DNA fragments were separated into each size. Based on its migration time, estimation of fragment size of the several restriction digests was performed without internal stan dards. And an enzyme-hydrolysed glucomannan with low molecular weight was also applied to separate oligonucleotides in CE. Highly reproducible separation was achieved by refilling the capillay with fresh glucomannan solution in each analytical cycle.
We concluded that the glucomannan is a good alternative to the conventional sieving additives for CE, such as polyacrylamide and hydroxypropylmethylcellulose, and this method would be applied to molecular cloning of a gene, separation and analysis of DNA fragments generated by restriction enzyme digestion or produced by polymerase chain reaction.
