Electrophoretic extraction and analysis of DNA from chitosan or poly-l-lysine-coated alginate beads |
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Authors: | Douglas Quong Ronald J Neufeld |
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Institution: | (1) Department of Chemical Engineering, McGill University, 3480 University Street, H3A 2A7 Montréal, Québec, Canada;(2) Present address: 3M Canada, P.O. Box 5757, N6A 4T1 London, Ontario, Canada;(3) Present address: Department of Chemical Engineering, Queen’s University, K7L 3N6 Kingston, Ontario, Canada |
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Abstract: | Alginate beads containing entrapped DNA were produced using both external and internal calcium sources, and coated with chitosan
or poly-l-lysine membranes. The beads were assayed with DNase nuclease to determine formulation conditions offering the highest level
of DNA protection fromnucleic acid hydrolysis, simulating gastrointestinal exposure. A method was developed to extract and
assay intracapsular DNA through a modified agarose electrophoresis system. Both external and internally gelled beads were
permeable to DNase (Mw=31 kDa), indicated by the absence of DNA after nuclease exposure. At low levels of DNase exposure, coated high guluronic
content alginate beads offered a higher level of DNA protection compared with coated beads with low guluronic alginate. No
apparent correlation was found with chitosan membrane molecular weight and degree of deacetylation; however, increasing poly-l-lysine molecular weight appeared to increase DNase exclusion from beads. At elevated levels of DNase exposure, DNA hydrolysis
was evident within all coated beads with the exception of those coated with the highest molecular weight poly-l-lysine (Mw=197.1 kDa), which provided almost total nuclease protection. Optimal combination then for DNA protection from nucleases is
a high guluronic alginate core, coated with high molecular weight poly-l-lysine. |
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Keywords: | DNA chitosan l-lysine" target="_blank">poly-l-lysine alginate electrophoresis |
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